Inhibitors of lysine specific demethylase-1

ABSTRACT

The present invention relates generally to compositions and methods for treating cancer and neoplastic disease. Provided herein are substituted heterocyclic derivative compounds and pharmaceutical compositions comprising said compounds. The subject compounds and compositions are useful for inhibition of lysine specific demethylase-1. Furthermore, the subject compounds and compositions are useful for the treatment of cancer, such as prostate cancer, breast cancer, bladder cancer, lung cancer and/or melanoma and the like.

CROSS-REFERENCE

This application claims the benefit of U.S. Provisional Application62/018,365, filed Jun. 27, 2014, which is incorporated herein byreference in its entirety.

BACKGROUND

A need exists in the art for an effective treatment of cancer andneoplastic disease.

BRIEF SUMMARY OF THE INVENTION

Provided herein are substituted heterocyclic derivative compounds andpharmaceutical compositions comprising said compounds. The subjectcompounds and compositions are useful for inhibition lysine specificdemethylase-1 (LSD-1). Furthermore, the subject compounds andcompositions are useful for the treatment of cancer, such as prostatecancer, breast cancer, bladder cancer, lung cancer and/or melanoma andthe like. The substituted heterocyclic derivative compounds describedherein are based upon a central heterocyclic ring system, such as apyridinone or pyrazinone, or the like. Said central heterocyclic ringsystem is further substituted with additional substituents, such as, forexample, a 4-cyanophenyl group and a heterocyclyl group.

One embodiment provides a compound having the structure of Formula (I),or a pharmaceutically acceptable salt thereof,

wherein,

-   -   A is N or C—R, wherein R is hydrogen or optionally substituted        alkyl;    -   W¹ and W² are independently chosen from N, C—H, or C—F;    -   X is hydrogen, halogen, optionally substituted alkyl, optionally        substituted cycloalkylalkyl, optionally substituted        heterocyclylalkyl, optionally substituted aralkyl, optionally        substituted heteroarylalkyl, optionally substituted alkyne,        optionally substituted carbocyclylalkynyl, optionally        substituted aryl, or optionally substituted heteroaryl;    -   Y is optionally substituted alkyl, optionally substituted        cycloalkylalkyl, heterocyclylalkyl or aralkyl; and    -   Z is an optionally substituted group chosen from N-heterocyclyl,        —O— heterocyclylalkyl, —N(H)-heterocyclylalkyl, or        —N(Me)-heterocyclylalkyl.

One embodiment provides a pharmaceutical composition comprising acompound of Formula (I), or a pharmaceutically acceptable salt thereof,and a pharmaceutically acceptable excipient.

One embodiment provides a method of regulating gene transcription in acell comprising inhibiting lysine-specific demethylase 1 activity byexposing the lysine-specific demethylase 1 enzyme to a compound ofFormula (I).

One embodiment provides a method of treating cancer in a patient in needthereof, comprising administering to the patient a compound of Formula(I), or a pharmaceutically acceptable salt thereof.

INCORPORATION BY REFERENCE

All publications, patents, and patent applications mentioned in thisspecification are herein incorporated by reference to the same extent asif each individual publication, patent, or patent application wasspecifically and individually indicated to be incorporated by reference.

DETAILED DESCRIPTION OF THE INVENTION

As used herein and in the appended claims, the singular forms “a,”“and,” and “the” include plural referents unless the context clearlydictates otherwise. Thus, for example, reference to “an agent” includesa plurality of such agents, and reference to “the cell” includesreference to one or more cells (or to a plurality of cells) andequivalents thereof known to those skilled in the art, and so forth.When ranges are used herein for physical properties, such as molecularweight, or chemical properties, such as chemical formulae, allcombinations and subcombinations of ranges and specific embodimentstherein are intended to be included. The term “about” when referring toa number or a numerical range means that the number or numerical rangereferred to is an approximation within experimental variability (orwithin statistical experimental error), and thus the number or numericalrange may vary between 1% and 15% of the stated number or numericalrange. The term “comprising” (and related terms such as “comprise” or“comprises” or “having” or “including”) is not intended to exclude thatin other certain embodiments, for example, an embodiment of anycomposition of matter, composition, method, or process, or the like,described herein, may “consist of” or “consist essentially of” thedescribed features.

DEFINITIONS

As used in the specification and appended claims, unless specified tothe contrary, the following terms have the meaning indicated below.

“Amino” refers to the —NH₂ radical.

“Cyano” refers to the —CN radical.

“Nitro” refers to the —NO₂ radical.

“Oxa” refers to the —O— radical.

“Oxo” refers to the ═O radical.

“Thioxo” refers to the ═S radical.

“Imino” refers to the ═N—H radical.

“Oximo” refers to the ═N—OH radical.

“Hydrazino” refers to the ═N—NH₂ radical.

“Alkyl” refers to a straight or branched hydrocarbon chain radicalconsisting solely of carbon and hydrogen atoms, containing nounsaturation, having from one to fifteen carbon atoms (e.g., C₁-C₁₅alkyl). In certain embodiments, an alkyl comprises one to thirteencarbon atoms (e.g., C₁-C₁₃ alkyl). In certain embodiments, an alkylcomprises one to eight carbon atoms (e.g., C₁-C₈ alkyl). In otherembodiments, an alkyl comprises one to five carbon atoms (e.g., C₁-C₅alkyl). In other embodiments, an alkyl comprises one to four carbonatoms (e.g., C₁-C₄ alkyl). In other embodiments, an alkyl comprises oneto three carbon atoms (e.g., C₁-C₃ alkyl). In other embodiments, analkyl comprises one to two carbon atoms (e.g., C₁-C₂ alkyl). In otherembodiments, an alkyl comprises one carbon atom (e.g., C₁ alkyl). Inother embodiments, an alkyl comprises five to fifteen carbon atoms(e.g., C₅-C₁₅ alkyl). In other embodiments, an alkyl comprises five toeight carbon atoms (e.g., C₅-C₈ alkyl). In other embodiments, an alkylcomprises two to five carbon atoms (e.g., C₂-C₅ alkyl). In otherembodiments, an alkyl comprises three to five carbon atoms (e.g., C₃-C₅alkyl). In other embodiments, the alkyl group is selected from methyl,ethyl, 1-propyl (n-propyl), 1-methylethyl (iso-propyl), 1-butyl(n-butyl), 1-methylpropyl (sec-butyl), 2-methylpropyl (iso-butyl),1,1-dimethylethyl (tert-butyl), 1-pentyl (n-pentyl). The alkyl isattached to the rest of the molecule by a single bond. Unless statedotherwise specifically in the specification, an alkyl group isoptionally substituted by one or more of the following substituents:halo, cyano, nitro, oxo, thioxo, imino, oximo, trimethylsilanyl,—OR^(a), —SR^(a), —OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a),—C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a), —OC(O)— N (R^(a))₂,—N(R^(a))C(O)R^(a), —N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2),—S(O)_(t)OR^(a) (where t is 1 or 2), —S(O)_(t)R^(a) (where t is 1 or 2)and —S(O)_(t)N(R^(a))₂ (where t is 1 or 2) where each R^(a) isindependently hydrogen, alkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, carbocyclyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), carbocyclylalkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl),heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), heterocyclylalkyl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl), orheteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy,or trifluoromethyl).

“Alkoxy” refers to a radical bonded through an oxygen atom of theformula —O-alkyl, where alkyl is an alkyl chain as defined above.

“Alkenyl” refers to a straight or branched hydrocarbon chain radicalgroup consisting solely of carbon and hydrogen atoms, containing atleast one carbon-carbon double bond, and having from two to twelvecarbon atoms. In certain embodiments, an alkenyl comprises two to eightcarbon atoms. In other embodiments, an alkenyl comprises two to fourcarbon atoms. The alkenyl is attached to the rest of the molecule by asingle bond, for example, ethenyl (i.e., vinyl), prop-1-enyl (i.e.,allyl), but-1-enyl, pent-1-enyl, penta-1,4-dienyl, and the like. Unlessstated otherwise specifically in the specification, an alkenyl group isoptionally substituted by one or more of the following substituents:halo, cyano, nitro, oxo, thioxo, imino, oximo, trimethylsilanyl,—OR^(a), —SR^(a), —OC(O)—R^(a), —N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a),—C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a), —OC(O)— N (R^(a))₂,—N(R^(a))C(O)R^(a), —N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2),—S(O)_(t)OR^(a) (where t is 1 or 2), —S(O)_(t)R^(a) (where t is 1 or 2)and —S(O)_(t)N(R^(a))₂ (where t is 1 or 2) where each R^(a) isindependently hydrogen, alkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, carbocyclyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), carbocyclylalkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl),heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), heterocyclylalkyl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl), orheteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy,or trifluoromethyl).

“Alkynyl” refers to a straight or branched hydrocarbon chain radicalgroup consisting solely of carbon and hydrogen atoms, containing atleast one carbon-carbon triple bond, having from two to twelve carbonatoms. In certain embodiments, an alkynyl comprises two to eight carbonatoms. In other embodiments, an alkynyl comprises two to six carbonatoms. In other embodiments, an alkynyl comprises two to four carbonatoms. The alkynyl is attached to the rest of the molecule by a singlebond, for example, ethynyl, propynyl, butyryl, pentynyl, hexynyl, andthe like. Unless stated otherwise specifically in the specification, analkynyl group is optionally substituted by one or more of the followingsubstituents: halo, cyano, nitro, oxo, thioxo, imino, oximo,trimethylsilanyl, —OR^(a), —SR^(a), —OC(O)—R^(a), —N(R^(a))₂,—C(O)R^(a), —C(O)OR^(a), —C(O)N(R^(a))₂, —N(R^(a))C(O)OR^(a), —OC(O)— N(R^(a))₂, —N(R^(a))C(O)R^(a), —N(R^(a))S(O)_(t)R^(a) (where t is 1 or2), —S(O)_(t)OR^(a) (where t is 1 or 2), —S(O)_(t)R^(a) (where t is 1 or2) and —S(O)_(t)N(R^(a))₂ (where t is 1 or 2) where each R^(a) isindependently hydrogen, alkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, carbocyclyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), carbocyclylalkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl),heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), heterocyclylalkyl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl), orheteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy,or trifluoromethyl).

“Alkylene” or “alkylene chain” refers to a straight or branched divalenthydrocarbon chain linking the rest of the molecule to a radical group,consisting solely of carbon and hydrogen, containing no unsaturation andhaving from one to twelve carbon atoms, for example, methylene,ethylene, propylene, n-butylene, and the like. The alkylene chain isattached to the rest of the molecule through a single bond and to theradical group through a single bond. The points of attachment of thealkylene chain to the rest of the molecule and to the radical group canbe through one carbon in the alkylene chain or through any two carbonswithin the chain. In certain embodiments, an alkylene comprises one toeight carbon atoms (e.g., C₁-C₈ alkylene). In other embodiments, analkylene comprises one to five carbon atoms (e.g., C₁-C₅ alkylene). Inother embodiments, an alkylene comprises one to four carbon atoms (e.g.,C₁-C₄ alkylene). In other embodiments, an alkylene comprises one tothree carbon atoms (e.g., C₁-C₃ alkylene). In other embodiments, analkylene comprises one to two carbon atoms (e.g., C₁-C₂ alkylene). Inother embodiments, an alkylene comprises one carbon atom (e.g., C₁alkylene). In other embodiments, an alkylene comprises five to eightcarbon atoms (e.g., C₅-C₈ alkylene). In other embodiments, an alkylenecomprises two to five carbon atoms (e.g., C₂-C₅ alkylene). In otherembodiments, an alkylene comprises three to five carbon atoms (e.g.,C₃-C₅ alkylene). Unless stated otherwise specifically in thespecification, an alkylene chain is optionally substituted by one ormore of the following substituents: halo, cyano, nitro, oxo, thioxo,imino, oximo, trimethylsilanyl, —OR^(a), —SR^(a), —OC(O)—R^(a),—N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a), —C(O)N(R^(a))₂,—N(R^(a))C(O)OR^(a), —OC(O)— N (R^(a))₂, —N(R^(a))C(O)R^(a),—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —S(O)_(t)OR^(a) (where t is1 or 2), —S(O)_(t)R^(a) (where t is 1 or 2) and —S(O)_(t)N(R^(a))₂(where t is 1 or 2) where each R^(a) is independently hydrogen, alkyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), fluoroalkyl, carbocyclyl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), carbocyclylalkyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), aryl (optionally substituted with halogen, hydroxy,methoxy, or trifluoromethyl), aralkyl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), heterocyclyl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl),heterocyclylalkyl (optionally substituted with halogen, hydroxy,methoxy, or trifluoromethyl), heteroaryl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), or heteroarylalkyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl).

“Alkynylene” or “alkynylene chain” refers to a straight or brancheddivalent hydrocarbon chain linking the rest of the molecule to a radicalgroup, consisting solely of carbon and hydrogen, containing at least onecarbon-carbon triple bond, and having from two to twelve carbon atoms.The alkynylene chain is attached to the rest of the molecule through asingle bond and to the radical group through a single bond. In certainembodiments, an alkynylene comprises two to eight carbon atoms (e.g.,C₂-C₈ alkynylene). In other embodiments, an alkynylene comprises two tofive carbon atoms (e.g., C₂-C₅ alkynylene). In other embodiments, analkynylene comprises two to four carbon atoms (e.g., C₂-C₄ alkynylene).In other embodiments, an alkynylene comprises two to three carbon atoms(e.g., C₂-C₃ alkynylene). In other embodiments, an alkynylene comprisestwo carbon atom (e.g., C₂ alkylene). In other embodiments, an alkynylenecomprises five to eight carbon atoms (e.g., C₅-C₈ alkynylene). In otherembodiments, an alkynylene comprises three to five carbon atoms (e.g.,C₃-C₅ alkynylene). Unless stated otherwise specifically in thespecification, an alkynylene chain is optionally substituted by one ormore of the following substituents: halo, cyano, nitro, oxo, thioxo,imino, oximo, trimethylsilanyl, —OR^(a), —SR^(a), —OC(O)—R^(a),—N(R^(a))₂, —C(O)R^(a), —C(O)OR^(a), —C(O)N(R^(a))₂,—N(R^(a))C(O)OR^(a), —OC(O)— N (R^(a))₂, —N(R^(a))C(O)R^(a),—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —S(O)_(t)OR^(a) (where t is1 or 2), —S(O)_(t)R^(a) (where t is 1 or 2) and —S(O)_(t)N(R^(a))₂(where t is 1 or 2) where each R^(a) is independently hydrogen, alkyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), fluoroalkyl, carbocyclyl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), carbocyclylalkyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), aryl (optionally substituted with halogen, hydroxy,methoxy, or trifluoromethyl), aralkyl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), heterocyclyl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl),heterocyclylalkyl (optionally substituted with halogen, hydroxy,methoxy, or trifluoromethyl), heteroaryl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), or heteroarylalkyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl).

“Aryl” refers to a radical derived from an aromatic monocyclic ormulticyclic hydrocarbon ring system by removing a hydrogen atom from aring carbon atom. The aromatic monocyclic or multicyclic hydrocarbonring system contains only hydrogen and carbon from five to eighteencarbon atoms, where at least one of the rings in the ring system isfully unsaturated, i.e., it contains a cyclic, delocalized (4n+2)π-electron system in accordance with the Hückel theory. The ring systemfrom which aryl groups are derived include, but are not limited to,groups such as benzene, fluorene, indane, indene, tetralin andnaphthalene. Unless stated otherwise specifically in the specification,the term “aryl” or the prefix “ar-” (such as in “aralkyl”) is meant toinclude aryl radicals optionally substituted by one or more substituentsindependently selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl,cyano, nitro, optionally substituted aryl, optionally substitutedaralkyl, optionally substituted aralkenyl, optionally substitutedaralkynyl, optionally substituted carbocyclyl, optionally substitutedcarbocyclylalkyl, optionally substituted heterocyclyl, optionallysubstituted heterocyclylalkyl, optionally substituted heteroaryl,optionally substituted heteroarylalkyl, —R^(b)—OR^(a),—R^(b)—OC(O)—R^(a), —R^(b)—OC(O)—OR^(a), —R^(b)—OC(O)—N(R^(a))₂,—R^(b)—N(R^(a))₂, —R^(b)—C(O)R^(a), —R^(b)—C(O)OR^(a),—R^(b)—C(O)N(R^(a))₂, —R^(b)—O—R^(c)—C(O)N(R^(a))₂,—R^(b)—N(R^(a))C(O)OR^(a), —R^(b)—N(R^(a))C(O)R^(a),—R^(b)—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —R^(b)—S(O)_(t)R^(a)(where t is 1 or 2), —R^(b)—S(O)_(t)OR^(a) (where t is 1 or 2) and—R^(b)—S(O)_(t)N(R^(a))₂ (where t is 1 or 2), where each R^(a) isindependently hydrogen, alkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, cycloalkyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), cycloalkylalkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl),heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), heterocyclylalkyl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl), orheteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy,or trifluoromethyl), each R^(b) is independently a direct bond or astraight or branched alkylene or alkenylene chain, and R^(c) is astraight or branched alkylene or alkenylene chain, and where each of theabove substituents is unsubstituted unless otherwise indicated.

“Aralkyl” refers to a radical of the formula —R^(c)-aryl where R^(c) isan alkylene chain as defined above, for example, methylene, ethylene,and the like. The alkylene chain part of the aralkyl radical isoptionally substituted as described above for an alkylene chain. Thearyl part of the aralkyl radical is optionally substituted as describedabove for an aryl group.

“Aralkenyl” refers to a radical of the formula —R^(d)-aryl where R^(d)is an alkenylene chain as defined above. The aryl part of the aralkenylradical is optionally substituted as described above for an aryl group.The alkenylene chain part of the aralkenyl radical is optionallysubstituted as defined above for an alkenylene group.

“Aralkynyl” refers to a radical of the formula —R^(e)-aryl, where R^(e)is an alkynylene chain as defined above. The aryl part of the aralkynylradical is optionally substituted as described above for an aryl group.The alkynylene chain part of the aralkynyl radical is optionallysubstituted as defined above for an alkynylene chain.

“Aralkoxy” refers to a radical bonded through an oxygen atom of theformula —O—R^(c)-aryl where R^(c) is an alkylene chain as defined above,for example, methylene, ethylene, and the like. The alkylene chain partof the aralkyl radical is optionally substituted as described above foran alkylene chain. The aryl part of the aralkyl radical is optionallysubstituted as described above for an aryl group.

“Carbocyclyl” refers to a stable non-aromatic monocyclic or polycyclichydrocarbon radical consisting solely of carbon and hydrogen atoms,which includes fused or bridged ring systems, having from three tofifteen carbon atoms. In certain embodiments, a carbocyclyl comprisesthree to ten carbon atoms. In other embodiments, a carbocyclyl comprisesfive to seven carbon atoms. The carbocyclyl is attached to the rest ofthe molecule by a single bond. Carbocyclyl may be saturated, (i.e.,containing single C—C bonds only) or unsaturated (i.e., containing oneor more double bonds or triple bonds.) A fully saturated carbocyclylradical is also referred to as “cycloalkyl.” Examples of monocycliccycloalkyls include, e.g., cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl, and cyclooctyl. An unsaturated carbocyclyl isalso referred to as “cycloalkenyl.” Examples of monocyclic cycloalkenylsinclude, e.g., cyclopentenyl, cyclohexenyl, cycloheptenyl, andcyclooctenyl. Polycyclic carbocyclyl radicals include, for example,adamantyl, norbornyl (i.e., bicyclo[2.2.1]heptanyl), norbornenyl,decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. Unlessotherwise stated specifically in the specification, the term“carbocyclyl” is meant to include carbocyclyl radicals that areoptionally substituted by one or more substituents independentlyselected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl, oxo, thioxo,cyano, nitro, optionally substituted aryl, optionally substitutedaralkyl, optionally substituted aralkenyl, optionally substitutedaralkynyl, optionally substituted carbocyclyl, optionally substitutedcarbocyclylalkyl, optionally substituted heterocyclyl, optionallysubstituted heterocyclylalkyl, optionally substituted heteroaryl,optionally substituted heteroarylalkyl, —R^(b)—OR^(a),—R^(b)—OC(O)—R^(a), —R^(b)—OC(O)—OR^(a), —R^(b)—OC(O)—N(R^(a))₂,—R^(b)—N(R^(a))₂, —R^(b)—C(O)R^(a), —R^(b)—C(O)OR^(a),—R^(b)—C(O)N(R^(a))₂, —R^(b)—O—R^(c)—C(O)N(R^(a))₂,—R^(b)—N(R^(a))C(O)OR^(a), —R^(b)—N(R^(a))C(O)R^(a), —_(R)^(b)—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —_(R) ^(b)—S(O)_(t)R^(a)(where t is 1 or 2), —R^(b)—S(O)_(t)OR^(a) (where t is 1 or 2) and—R^(b)—S(O)_(t)N(R^(a))₂ (where t is 1 or 2), where each R^(a) isindependently hydrogen, alkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, cycloalkyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), cycloalkylalkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl),heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), heterocyclylalkyl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl), orheteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy,or trifluoromethyl), each R^(b) is independently a direct bond or astraight or branched alkylene or alkenylene chain, and R^(c) is astraight or branched alkylene or alkenylene chain, and where each of theabove substituents is unsubstituted unless otherwise indicated.

“Carbocyclylalkyl” refers to a radical of the formula —R^(c)-carbocyclylwhere R^(c) is an alkylene chain as defined above. The alkylene chainand the carbocyclyl radical is optionally substituted as defined above.

“Carbocyclylalkynyl” refers to a radical of the formula—R^(c)-carbocyclyl where R^(c) is an alkynylene chain as defined above.The alkynylene chain and the carbocyclyl radical is optionallysubstituted as defined above.

“Carbocyclylalkoxy” refers to a radical bonded through an oxygen atom ofthe formula —O—R^(c)-carbocyclyl where R^(c) is an alkylene chain asdefined above. The alkylene chain and the carbocyclyl radical isoptionally substituted as defined above.

As used herein, “carboxylic acid bioisostere” refers to a functionalgroup or moiety that exhibits similar physical, biological and/orchemical properties as a carboxylic acid moiety. Examples of carboxylicacid bioisosteres include, but are not limited to,

and the like.

“Halo” or “halogen” refers to bromo, chloro, fluoro or iodosubstituents.

“Fluoroalkyl” refers to an alkyl radical, as defined above, that issubstituted by one or more fluoro radicals, as defined above, forexample, trifluoromethyl, difluoromethyl, fluoromethyl,2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, and the like. Thealkyl part of the fluoroalkyl radical may be optionally substituted asdefined above for an alkyl group.

“Heterocyclyl” refers to a stable 3- to 18-membered non-aromatic ringradical that comprises two to twelve carbon atoms and from one to sixheteroatoms selected from nitrogen, oxygen and sulfur. Unless statedotherwise specifically in the specification, the heterocyclyl radical isa monocyclic, bicyclic, tricyclic or tetracyclic ring system, which mayinclude fused or bridged ring systems. The heteroatoms in theheterocyclyl radical may be optionally oxidized. One or more nitrogenatoms, if present, are optionally quaternized. The heterocyclyl radicalis partially or fully saturated. The heterocyclyl may be attached to therest of the molecule through any atom of the ring(s). Examples of suchheterocyclyl radicals include, but are not limited to, dioxolanyl,thienyl[1,3]dithianyl, decahydroisoquinolyl, imidazolinyl,imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl,octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl,2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl,piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrazolidinyl, quinuclidinyl,thiazolidinyl, tetrahydrofuryl, trithianyl, tetrahydropyranyl,thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and1,1-dioxo-thiomorpholinyl. Unless stated otherwise specifically in thespecification, the term “heterocyclyl” is meant to include heterocyclylradicals as defined above that are optionally substituted by one or moresubstituents selected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl,oxo, thioxo, cyano, nitro, optionally substituted aryl, optionallysubstituted aralkyl, optionally substituted aralkenyl, optionallysubstituted aralkynyl, optionally substituted carbocyclyl, optionallysubstituted carbocyclylalkyl, optionally substituted heterocyclyl,optionally substituted heterocyclylalkyl, optionally substitutedheteroaryl, optionally substituted heteroarylalkyl, —R^(b)—OR^(a),—R^(b)—OC(O)—R^(a), —R^(b)—OC(O)—OR^(a), —R^(b)—OC(O)—N(R^(a))₂,—R^(b)—N(R^(a))₂, —R^(b)—C(O)R^(a), —R^(b)—C(O)OR^(a),—R^(b)—C(O)N(R^(a))₂, —R^(b)—O—R^(c)—C(O)N(R^(a))₂,—R^(b)—N(R^(a))C(O)OR^(a), —R^(b)—N(R^(a))C(O)R^(a),—R^(b)—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —R^(b)—S(O)_(t)R^(a)(where t is 1 or 2), —R^(b)—S(O)_(t)OR^(a) (where t is 1 or 2) and—R^(b)—S(O)_(t)N(R^(a))₂ (where t is 1 or 2), where each R^(a) isindependently hydrogen, alkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, cycloalkyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), cycloalkylalkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl),heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), heterocyclylalkyl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl), orheteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy,or trifluoromethyl), each R^(b) is independently a direct bond or astraight or branched alkylene or alkenylene chain, and R^(c) is astraight or branched alkylene or alkenylene chain, and where each of theabove substituents is unsubstituted unless otherwise indicated.

“N-heterocyclyl” or “N-attached heterocyclyl” refers to a heterocyclylradical as defined above containing at least one nitrogen and where thepoint of attachment of the heterocyclyl radical to the rest of themolecule is through a nitrogen atom in the heterocyclyl radical. AnN-heterocyclyl radical is optionally substituted as described above forheterocyclyl radicals. Examples of such N-heterocyclyl radicals include,but are not limited to, 1-morpholinyl, 1-piperidinyl, 1-piperazinyl,1-pyrrolidinyl, pyrazolidinyl, imidazolinyl, and imidazolidinyl.

“C-heterocyclyl” or “C-attached heterocyclyl” refers to a heterocyclylradical as defined above containing at least one heteroatom and wherethe point of attachment of the heterocyclyl radical to the rest of themolecule is through a carbon atom in the heterocyclyl radical. AC-heterocyclyl radical is optionally substituted as described above forheterocyclyl radicals. Examples of such C-heterocyclyl radicals include,but are not limited to, 2-morpholinyl, 2- or 3- or 4-piperidinyl,2-piperazinyl, 2- or 3-pyrrolidinyl, and the like.

“Heterocyclylalkyl” refers to a radical of the formula—R^(c)-heterocyclyl where R^(c) is an alkylene chain as defined above.If the heterocyclyl is a nitrogen-containing heterocyclyl, theheterocyclyl is optionally attached to the alkyl radical at the nitrogenatom. The alkylene chain of the heterocyclylalkyl radical is optionallysubstituted as defined above for an alkylene chain. The heterocyclylpart of the heterocyclylalkyl radical is optionally substituted asdefined above for a heterocyclyl group.

“Heterocyclylalkoxy” refers to a radical bonded through an oxygen atomof the formula —O—R^(c)-heterocyclyl where R^(c) is an alkylene chain asdefined above. If the heterocyclyl is a nitrogen-containingheterocyclyl, the heterocyclyl is optionally attached to the alkylradical at the nitrogen atom. The alkylene chain of theheterocyclylalkoxy radical is optionally substituted as defined abovefor an alkylene chain. The heterocyclyl part of the heterocyclylalkoxyradical is optionally substituted as defined above for a heterocyclylgroup.

“Heteroaryl” refers to a radical derived from a 3- to 18-memberedaromatic ring radical that comprises two to seventeen carbon atoms andfrom one to six heteroatoms selected from nitrogen, oxygen and sulfur.As used herein, the heteroaryl radical may be a monocyclic, bicyclic,tricyclic or tetracyclic ring system, wherein at least one of the ringsin the ring system is fully unsaturated, i.e., it contains a cyclic,delocalized (4n+2) π-electron system in accordance with the Hückeltheory. Heteroaryl includes fused or bridged ring systems. Theheteroatom(s) in the heteroaryl radical is optionally oxidized. One ormore nitrogen atoms, if present, are optionally quaternized. Theheteroaryl is attached to the rest of the molecule through any atom ofthe ring(s). Examples of heteroaryls include, but are not limited to,azepinyl, acridinyl, benzimidazolyl, benzindolyl, 1,3-benzodioxolyl,benzofuranyl, benzooxazolyl, benzo[d]thiazolyl, benzothiadiazolyl,benzo[b][1,4]dioxepinyl, benzo[b][1,4]oxazinyl, 1,4-benzodioxanyl,benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl,benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl(benzothiophenyl), benzothieno[3,2-d]pyrimidinyl, benzotriazolyl,benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl,cyclopenta[d]pyrimidinyl,6,7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d]pyrimidinyl,5,6-dihydrobenzo[h]quinazolinyl, 5,6-dihydrobenzo[h]cinnolinyl,6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridazinyl, dibenzofuranyl,dibenzothiophenyl, furanyl, furanonyl, furo[3,2-c]pyridinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyrimidinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyridazinyl,5,6,7,8,9,10-hexahydrocycloocta[d]pyridinyl, isothiazolyl, imidazolyl,indazolyl, indolyl, indazolyl, isoindolyl, indolinyl, isoindolinyl,isoquinolyl, indolizinyl, isoxazolyl,5,8-methano-5,6,7,8-tetrahydroquinazolinyl, naphthyridinyl,1,6-naphthyridinonyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl,5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl, 1-phenyl-1H-pyrrolyl,phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl,purinyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl, pyridinyl,pyrido[3,2-d]pyrimidinyl, pyrido[3,4-d]pyrimidinyl, pyrazinyl,pyrimidinyl, pyridazinyl, pyrrolyl, quinazolinyl, quinoxalinyl,quinolinyl, isoquinolinyl, tetrahydroquinolinyl,5,6,7,8-tetrahydroquinazolinyl,5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl,6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d]pyrimidinyl,5,6,7,8-tetrahydropyrido[4,5-c]pyridazinyl, thiazolyl, thiadiazolyl,triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl,thieno[3,2-d]pyrimidinyl, thieno[2,3-c]pridinyl, and thiophenyl (i.e.thienyl). Unless stated otherwise specifically in the specification, theterm “heteroaryl” is meant to include heteroaryl radicals as definedabove which are optionally substituted by one or more substituentsselected from alkyl, alkenyl, alkynyl, halo, fluoroalkyl, haloalkenyl,haloalkynyl, oxo, thioxo, cyano, nitro, optionally substituted aryl,optionally substituted aralkyl, optionally substituted aralkenyl,optionally substituted aralkynyl, optionally substituted carbocyclyl,optionally substituted carbocyclylalkyl, optionally substitutedheterocyclyl, optionally substituted heterocyclylalkyl, optionallysubstituted heteroaryl, optionally substituted heteroarylalkyl,—R^(b)—OR^(a), —R^(b)—OC(O)—R^(a), —R^(b)—OC(O)—OR^(a),—R^(b)—OC(O)—N(R^(a))₂, —R^(b)—N(R^(a))², —R^(b)—C(O)R^(a),—R^(b)—C(O)OR^(a), —R^(b)—C(O)N(R^(a))₂, —R^(b)—O—R^(c)—C(O)N(R^(a))₂,—R^(b)—N(R^(a))C(O)OR^(a), —R^(b)—N(R^(a))C(O)R^(a),—R^(b)—N(R^(a))S(O)_(t)R^(a) (where t is 1 or 2), —R^(b)—S(O)_(t)R^(a)(where t is 1 or 2), —R^(b)—S(O)_(t)OR^(a) (where t is 1 or 2) and—R^(b)—S(O)_(t)N(R^(a))₂ (where t is 1 or 2), where each R^(a) isindependently hydrogen, alkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, cycloalkyl(optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), cycloalkylalkyl (optionally substituted with halogen,hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl),heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, ortrifluoromethyl), heterocyclylalkyl (optionally substituted withhalogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionallysubstituted with halogen, hydroxy, methoxy, or trifluoromethyl), orheteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy,or trifluoromethyl), each R^(b) is independently a direct bond or astraight or branched alkylene or alkenylene chain, and R^(c) is astraight or branched alkylene or alkenylene chain, and where each of theabove substituents is unsubstituted unless otherwise indicated.

“N-heteroaryl” refers to a heteroaryl radical as defined abovecontaining at least one nitrogen and where the point of attachment ofthe heteroaryl radical to the rest of the molecule is through a nitrogenatom in the heteroaryl radical. An N-heteroaryl radical is optionallysubstituted as described above for heteroaryl radicals.

“C-heteroaryl” refers to a heteroaryl radical as defined above and wherethe point of attachment of the heteroaryl radical to the rest of themolecule is through a carbon atom in the heteroaryl radical. AC-heteroaryl radical is optionally substituted as described above forheteroaryl radicals.

“Heteroarylalkyl” refers to a radical of the formula —R^(c)-heteroaryl,where R^(c) is an alkylene chain as defined above. If the heteroaryl isa nitrogen-containing heteroaryl, the heteroaryl is optionally attachedto the alkyl radical at the nitrogen atom. The alkylene chain of theheteroarylalkyl radical is optionally substituted as defined above foran alkylene chain. The heteroaryl part of the heteroarylalkyl radical isoptionally substituted as defined above for a heteroaryl group.

“Heteroarylalkoxy” refers to a radical bonded through an oxygen atom ofthe formula —O—R^(c)-heteroaryl, where R^(c) is an alkylene chain asdefined above. If the heteroaryl is a nitrogen-containing heteroaryl,the heteroaryl is optionally attached to the alkyl radical at thenitrogen atom. The alkylene chain of the heteroarylalkoxy radical isoptionally substituted as defined above for an alkylene chain. Theheteroaryl part of the heteroarylalkoxy radical is optionallysubstituted as defined above for a heteroaryl group.

The compounds disclosed herein may contain one or more asymmetriccenters and may thus give rise to enantiomers, diastereomers, and otherstereoisomeric forms that may be defined, in terms of absolutestereochemistry, as (R)— or (S)—. Unless stated otherwise, it isintended that all stereoisomeric forms of the compounds disclosed hereinare contemplated by this disclosure. When the compounds described hereincontain alkene double bonds, and unless specified otherwise, it isintended that this disclosure includes both E and Z geometric isomers(e.g., cis or trans.) Likewise, all possible isomers, as well as theirracemic and optically pure forms, and all tautomeric forms are alsointended to be included. The term “geometric isomer” refers to E or Zgeometric isomers (e.g., cis or trans) of an alkene double bond. Theterm “positional isomer” refers to structural isomers around a centralring, such as ortho-, meta-, and para-isomers around a benzene ring.

A “tautomer” refers to a molecule wherein a proton shift from one atomof a molecule to another atom of the same molecule is possible. Thecompounds presented herein may, in certain embodiments, exist astautomers. In circumstances where tautomerization is possible, achemical equilibrium of the tautomers will exist. The exact ratio of thetautomers depends on several factors, including physical state,temperature, solvent, and pH. Some examples of tautomeric equilibriuminclude:

“Optional” or “optionally” means that a subsequently described event orcircumstance may or may not occur and that the description includesinstances when the event or circumstance occurs and instances in whichit does not. For example, “optionally substituted aryl” means that thearyl radical may or may not be substituted and that the descriptionincludes both substituted aryl radicals and aryl radicals having nosubstitution.

“Pharmaceutically acceptable salt” includes both acid and base additionsalts. A pharmaceutically acceptable salt of any one of the substitutedheterocyclic derivative compounds described herein is intended toencompass any and all pharmaceutically suitable salt forms. Preferredpharmaceutically acceptable salts of the compounds described herein arepharmaceutically acceptable acid addition salts and pharmaceuticallyacceptable base addition salts.

“Pharmaceutically acceptable acid addition salt” refers to those saltswhich retain the biological effectiveness and properties of the freebases, which are not biologically or otherwise undesirable, and whichare formed with inorganic acids such as hydrochloric acid, hydrobromicacid, sulfuric acid, nitric acid, phosphoric acid, hydroiodic acid,hydrofluoric acid, phosphorous acid, and the like. Also included aresalts that are formed with organic acids such as aliphatic mono- anddicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoicacids, alkanedioic acids, aromatic acids, aliphatic and, aromaticsulfonic acids, etc. and include, for example, acetic acid,trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid,oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid,tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid,salicylic acid, and the like. Exemplary salts thus include sulfates,pyrosulfates, bisulfates, sulfites, bisulfites, nitrates, phosphates,monohydrogenphosphates, dihydrogenphosphates, metaphosphates,pyrophosphates, chlorides, bromides, iodides, acetates,trifluoroacetates, propionates, caprylates, isobutyrates, oxalates,malonates, succinate suberates, sebacates, fumarates, maleates,mandelates, benzoates, chlorobenzoates, methylbenzoates,dinitrobenzoates, phthalates, benzenesulfonates, toluenesulfonates,phenylacetates, citrates, lactates, malates, tartrates,methanesulfonates, and the like. Also contemplated are salts of aminoacids, such as arginates, gluconates, and galacturonates (see, forexample, Berge S. M. et al., “Pharmaceutical Salts,” Journal ofPharmaceutical Science, 66:1-19 (1997), which is hereby incorporated byreference in its entirety). Acid addition salts of basic compounds maybe prepared by contacting the free base forms with a sufficient amountof the desired acid to produce the salt according to methods andtechniques with which a skilled artisan is familiar.

“Pharmaceutically acceptable base addition salt” refers to those saltsthat retain the biological effectiveness and properties of the freeacids, which are not biologically or otherwise undesirable. These saltsare prepared from addition of an inorganic base or an organic base tothe free acid. Pharmaceutically acceptable base addition salts may beformed with metals or amines, such as alkali and alkaline earth metalsor organic amines. Salts derived from inorganic bases include, but arenot limited to, sodium, potassium, lithium, ammonium, calcium,magnesium, iron, zinc, copper, manganese, aluminum salts and the like.Salts derived from organic bases include, but are not limited to, saltsof primary, secondary, and tertiary amines, substituted amines includingnaturally occurring substituted amines, cyclic amines and basic ionexchange resins, for example, isopropylamine, trimethylamine,diethylamine, triethylamine, tripropylamine, ethanolamine,diethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol,dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine,N,N-dibenzylethylenediamine, chloroprocaine, hydrabamine, choline,betaine, ethylenediamine, ethylenedianiline, N-methylglucamine,glucosamine, methylglucamine, theobromine, purines, piperazine,piperidine, N-ethylpiperidine, polyamine resins and the like. See Bergeet al., supra.

As used herein, “treatment” or “treating,” or “palliating” or“ameliorating” are used interchangeably herein. These terms refers to anapproach for obtaining beneficial or desired results including but notlimited to therapeutic benefit and/or a prophylactic benefit. By“therapeutic benefit” is meant eradication or amelioration of theunderlying disorder being treated. Also, a therapeutic benefit isachieved with the eradication or amelioration of one or more of thephysiological symptoms associated with the underlying disorder such thatan improvement is observed in the patient, notwithstanding that thepatient may still be afflicted with the underlying disorder. Forprophylactic benefit, the compositions may be administered to a patientat risk of developing a particular disease, or to a patient reportingone or more of the physiological symptoms of a disease, even though adiagnosis of this disease may not have been made.

“Prodrug” is meant to indicate a compound that may be converted underphysiological conditions or by solvolysis to a biologically activecompound described herein. Thus, the term “prodrug” refers to aprecursor of a biologically active compound that is pharmaceuticallyacceptable. A prodrug may be inactive when administered to a subject,but is converted in vivo to an active compound, for example, byhydrolysis. The prodrug compound often offers advantages of solubility,tissue compatibility or delayed release in a mammalian organism (see,e.g., Bundgard, H., Design of Prodrugs (1985), pp. 7-9, 21-24 (Elsevier,Amsterdam).

A discussion of prodrugs is provided in Higuchi, T., et al., “Pro-drugsas Novel Delivery Systems,” A.C.S. Symposium Series, Vol. 14, and inBioreversible Carriers in Drug Design, ed. Edward B. Roche, AmericanPharmaceutical Association and Pergamon Press, 1987, both of which areincorporated in full by reference herein.

The term “prodrug” is also meant to include any covalently bondedcarriers, which release the active compound in vivo when such prodrug isadministered to a mammalian subject. Prodrugs of an active compound, asdescribed herein, may be prepared by modifying functional groups presentin the active compound in such a way that the modifications are cleaved,either in routine manipulation or in vivo, to the parent activecompound. Prodrugs include compounds wherein a hydroxy, amino ormercapto group is bonded to any group that, when the prodrug of theactive compound is administered to a mammalian subject, cleaves to forma free hydroxy, free amino or free mercapto group, respectively.Examples of prodrugs include, but are not limited to, acetate, formateand benzoate derivatives of alcohol or amine functional groups in theactive compounds and the like.

Substituted Heterocyclic Derivative Compounds

Substituted heterocyclic derivative compounds are described herein thatare lysine specific demethylase-1 inhibitors. These compounds, andcompositions comprising these compounds, are useful for the treatment ofcancer and neoplastic disease. The compounds described herein are usefulfor treating prostate cancer, breast cancer, bladder cancer, lung cancerand/or melanoma and the like.

One embodiment provides a compound having the structure of Formula (I),or a pharmaceutically acceptable salt thereof,

wherein,

-   -   A is N or C—R, wherein R is hydrogen or optionally substituted        alkyl;    -   W¹ and W² are independently chosen from N, C—H, or C—F;    -   X is hydrogen, halogen, optionally substituted alkyl, optionally        substituted cycloalkylalkyl, optionally substituted        heterocyclylalkyl, optionally substituted aralkyl, optionally        substituted heteroarylalkyl, optionally substituted alkyne,        optionally substituted carbocyclylalkynyl, optionally        substituted aryl, or optionally substituted heteroaryl;    -   Y is optionally substituted alkyl, optionally substituted        cycloalkylalkyl, heterocyclylalkyl or aralkyl; and    -   Z is an optionally substituted group chosen from N-heterocyclyl,        —O— heterocyclylalkyl, —N(H)-heterocyclylalkyl, or        —N(Me)-heterocyclylalkyl.

Another embodiment provides the compound or pharmaceutically acceptablesalt of Formula (I), wherein W² is C—H. Another embodiment provides thecompound or pharmaceutically acceptable salt of Formula (I), wherein W¹is C—F. Another embodiment provides the compound or pharmaceuticallyacceptable salt of Formula (I), wherein W¹ is C—H. Another embodimentprovides the compound or pharmaceutically acceptable salt of Formula(I), wherein W¹ is N.

Another embodiment provides the compound or pharmaceutically acceptablesalt of Formula (I), wherein X is hydrogen. Another embodiment providesthe compound or pharmaceutically acceptable salt of Formula (I), whereinX is optionally substituted alkyl. Another embodiment provides thecompound or pharmaceutically acceptable salt of Formula (I), wherein Xis optionally substituted alkyne. Another embodiment provides thecompound or pharmaceutically acceptable salt of Formula (I), wherein Xis optionally substituted carbocyclylalkynyl. Another embodimentprovides the compound or pharmaceutically acceptable salt of Formula(I), wherein X is optionally substituted aryl, or optionally substitutedheteroaryl. Another embodiment provides the compound or pharmaceuticallyacceptable salt of Formula (I), wherein X is optionally substitutedaryl. Another embodiment provides the compound or pharmaceuticallyacceptable salt of Formula (I), wherein X is optionally substituted aryland the optionally substituted aryl is an optionally substituted phenyl.Another embodiment provides the compound or pharmaceutically acceptablesalt of Formula (I), wherein X is optionally substituted heteroaryl.Another embodiment provides the compound or pharmaceutically acceptablesalt of Formula (I), wherein X is optionally substituted heteroaryl andthe optionally substituted heteroaryl is chosen from an optionallysubstituted pyridinyl, optionally substituted pyrazolyl, or optionallysubstituted indazolyl.

Another embodiment provides the compound or pharmaceutically acceptablesalt of Formula (I), wherein Z is an optionally substituted—O-heterocyclylalkyl. Another embodiment provides the compound orpharmaceutically acceptable salt of Formula (I), wherein Z is anoptionally substituted —O-heterocyclylalkyl and the heterocyclylalkylgroup has the formula —R^(c)-heterocyclyl and the R^(c) is an optionallysubstituted C₁-C₃ alkylene chain. Another embodiment provides thecompound or pharmaceutically acceptable salt of Formula (I), wherein Zis an optionally substituted —O— heterocyclylalkyl and theheterocyclylalkyl group has the formula —R^(c)-heterocyclyl and theR^(c) is an optionally substituted C₁ alkylene chain. Another embodimentprovides the compound or pharmaceutically acceptable salt of Formula(I), wherein Z is an optionally substituted —O-heterocyclylalkyl and theheterocyclylalkyl group has the formula —R^(c)-heterocyclyl and theheterocyclyl is an optionally substituted nitrogen-containing 4-, 5-,6-, or 7-membered heterocyclyl. Another embodiment provides the compoundor pharmaceutically acceptable salt of Formula (I), wherein Z is anoptionally substituted —N(H)-heterocyclylalkyl. Another embodimentprovides the compound or pharmaceutically acceptable salt of Formula(I), wherein Z is an optionally substituted —N(H)-heterocyclylalkyl andthe heterocyclylalkyl group has the formula —R^(c)-heterocyclyl and theR^(c) is an optionally substituted C₁-C₃ alkylene chain. Anotherembodiment provides the compound or pharmaceutically acceptable salt ofFormula (I), wherein Z is an optionally substituted—N(H)-heterocyclylalkyl and the heterocyclylalkyl group has the formula—R^(c)-heterocyclyl and the R^(c) is an optionally substituted C₁alkylene chain. Another embodiment provides the compound orpharmaceutically acceptable salt of Formula (I), wherein Z is anoptionally substituted —N(H)-heterocyclylalkyl and the heterocyclylalkylgroup has the formula —R^(c)-heterocyclyl and the heterocyclyl is anoptionally substituted nitrogen-containing 4-, 5-, 6-, or 7-memberedheterocyclyl. Another embodiment provides the compound orpharmaceutically acceptable salt of Formula (I), wherein Z is anoptionally substituted —N(Me)-heterocyclylalkyl. Another embodimentprovides the compound or pharmaceutically acceptable salt of Formula(I), wherein Z is an optionally substituted —N(Me)-heterocyclylalkyl andthe heterocyclylalkyl group has the formula —R^(c)-heterocyclyl and theR^(c) is an optionally substituted C₁-C₃ alkylene chain. Anotherembodiment provides the compound or pharmaceutically acceptable salt ofFormula (I), wherein Z is an optionally substituted—N(Me)-heterocyclylalkyl and the heterocyclylalkyl group has the formula—R^(c)-heterocyclyl and the R^(c) is an optionally substituted C₁alkylene chain. Another embodiment provides the compound orpharmaceutically acceptable salt of Formula (I), wherein Z is anoptionally substituted —N(Me)-heterocyclylalkyl and theheterocyclylalkyl group has the formula —R^(c)-heterocyclyl and theheterocyclyl is an optionally substituted nitrogen-containing 4-, 5-,6-, or 7-membered heterocyclyl.

Another embodiment provides the compound or pharmaceutically acceptablesalt of Formula (I), wherein Z is an optionally substitutedN-heterocyclyl. Another embodiment provides the compound orpharmaceutically acceptable salt of Formula (I), wherein Z is anoptionally substituted N-heterocyclyl and the optionally substitutedN-heterocyclyl is a 4-, 5-, 6-, or 7-membered N-heterocyclyl. Anotherembodiment provides the compound or pharmaceutically acceptable salt ofFormula (I), wherein Z is an optionally substituted N-heterocyclyl andthe optionally substituted N-heterocyclyl is a 6-memberedN-heterocyclyl. Another embodiment provides the compound orpharmaceutically acceptable salt of Formula (I), wherein Z is anoptionally substituted N-heterocyclyl and the optionally substitutedN-heterocyclyl is an optionally substituted piperidine. Anotherembodiment provides the compound or pharmaceutically acceptable salt ofFormula (I), wherein Z is an optionally substituted N-heterocyclyl andthe optionally substituted piperidine is an optionally substituted4-aminopiperidine.

Another embodiment provides the compound or pharmaceutically acceptablesalt of Formula (I), wherein Y is optionally substitutedcycloalkylalkyl. Another embodiment provides the compound orpharmaceutically acceptable salt of Formula (I), wherein Y is optionallysubstituted heterocyclylalkyl. Another embodiment provides the compoundor pharmaceutically acceptable salt of Formula (I), wherein Y isoptionally substituted aralkyl. Another embodiment provides the compoundor pharmaceutically acceptable salt of Formula (I), wherein Y isoptionally substituted alkyl. Another embodiment provides the compoundor pharmaceutically acceptable salt of Formula (I), wherein Y isoptionally substituted alkyl and the optionally substituted alkyl is anoptionally substituted C₁-C₃ alkyl. Another embodiment provides thecompound or pharmaceutically acceptable salt of Formula (I), wherein Yis optionally substituted alkyl and the optionally substituted alkyl isan optionally substituted C₁ alkyl. Another embodiment provides thecompound or pharmaceutically acceptable salt of Formula (I), wherein Yis optionally substituted alkyl and the optionally substituted alkyl isa methyl group.

Another embodiment provides the compound or pharmaceutically acceptablesalt of Formula (I), wherein A is N. Another embodiment provides thecompound or pharmaceutically acceptable salt of Formula (I), wherein Ais C—R. Another embodiment provides the compound or pharmaceuticallyacceptable salt of Formula (I), wherein A is C—R, and R is hydrogen.Another embodiment provides the compound or pharmaceutically acceptablesalt of Formula (I), wherein A is C—R and R is optionally substitutedalkyl.

In some embodiments, the substituted heterocyclic derivative compounddescribed herein has the structure provided in Table 1.

TABLE 1 Chemical Synthesis Example Structure Name 1

4-[6-(4-aminopiperidin-1-yl)-4- benzyl-5-oxo-4,5-dihydropyrazin-2-yl]benzonitrile 2

4-[6-(4-aminopiperidin-1-yl)-4- [(4-methylphenyl)methyl]-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 3

4-[6-(4-aminopiperidin-1-yl)-4- [(2-fluorophenyl)methyl]-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 4

4-[6-(4-aminopiperidin-1-yl)-4-(3- chloro-benzyl)-5-oxo-4,5-dihydro-pyrazin-2-yl]-benzonitrile 5

4-[6-(4-amino-piperidin-1-yl)-4- methyl-5-oxo-4,5-dihydro-pyrazin-2-yl]-2-fluoro-benzonitrile 6

4-[6-(4-Amino-piperidin-1-yl)-4- ethyl-5-oxo-4,5-dihydro-pyrazin-2-yl]-2-fluoro-benzonitrile 7

4-[6-(4-amino-piperidin-1-yl)-4- cyclopropylmethyl-5-oxo-4,5-dihydro-pyrazin-2-yl]-2-fluoro- benzonitrile 8

4-[6-(4-aminopiperidin-1-yl)-4- [(3-fluorophenyl)methyl]-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 9

4-[6-(4-aminopiperidin-1-yl)-4- [(4-fluorophenyl)methyl]-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 10

4-[6-(4-aminopiperidin-1-yl)-4- [(3-methoxyphenyl)methyl]-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 11

4-[6-(4-aminopiperidin-1-yl)-4- [(4-methoxyphenyl)methyl]-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 12

4-[6-(4-aminopiperidin-1-yl)-5- oxo-4-[(1R)-1-phenylethyl]-4,5-dihydropyrazin-2-yl]benzonitrile 13

4-[6-(4-aminopiperidin-1-yl)-5- oxo-4-[(1S)-1-phenylethyl]-4,5-dihydropyrazin-2-yl]benzonitrile 14

4-[6-(4-amino-piperidin-1-yl)-5- oxo-4-(tetrahydro-furan-3-ylmethyl)-4,5-dihydro-pyrazin-2- yl]-2-fluoro-benzonitrile 15

4-[6-(4-amino-piperidin-1-yl)-5- oxo-4-(tetrahydro-pyran-4-ylmethyl)-4,5-dihydro-pyrazin-2- yl]-2-fluoro-benzonitrile 16

4-[6-(4-aminopiperidin-1-yl)-4- methyl-3-(4-methylphenyl)-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 17

4-[4-methyl-3-(4-methylphenyl)- 5-oxo-6-{[(3S)-pyrrolidin-3-ylmethyl]amino}-4,5- dihydropyrazin-2-yl]benzonitrile 18

4-[4-methyl-3-(4-methylphenyl)- 5-oxo-6-{[(3R)-pyrrolidin-3-ylmethyl]amino}-4,5- dihydropyrazin-2-yl]benzonitrile 19

5-[6-(4-aminopiperidin-1-yl)-4- methyl-3-(4-methylphenyl)-5-oxo-4,5-dihydropyrazin-2-yl]pyridine- 2-carbonitrile 20

4-{4-methyl-6-[4- (methylamino)piperidin-1-yl]-3-(4-methylphenyl)-5-oxo-4,5- dihydropyrazin-2-yl}benzonitrile 21

4-[6-(4-amino-4-methylpiperidin- 1-yl)-4-methyl-3-(4-methylphenyl)-5-oxo-4,5- dihydropyrazin-2-yl]benzonitrile 22

4-[6-(3-aminopiperidin-1-yl)-4- methyl-3-(4-methylphenyl)-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 23

4-[4-methyl-3-(4-methylphenyl)- 6-{octahydro-1H-pyrrolo[3,4-c]pyridin-5-yl}-5-oxo-4,5- dihydropyrazin-2-yl]benzonitrile 24

4-(6-{2,8-diazaspiro[4.5]decan-8- yl}-4-methyl-3-(4-methylphenyl)-5-oxo-4,5-dihydropyrazin-2- yl)benzonitrile 25

4-(6-{decahydropyrrolo[3,4- d]azepin-6-yl}-4-methyl-3-(4-methylphenyl)-5-oxo-4,5- dihydropyrazin-2-yl)benzonitrile 26

4-[6-(4-aminopiperidin-1-yl)-4- ethyl-3-(4-methylphenyl)-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 27

4-[6-(4-aminopiperidin-1-yl)-3-(4- ethylphenyl)-4-methyl-5-oxo-4,5-dihydropyrazin-2-yl]benzonitrile 28

4-[3-(4-ethylphenyl)-4-methyl-5- oxo-6-{[(3S)-pyrrolidin-3-ylmethyl]amino}-4,5- dihydropyrazin-2-yl]benzonitrile 29

4-[3-(4-ethylphenyl)-4-methyl-5- oxo-6-{[(3R)-pyrrolidin-3-ylmethyl]amino}-4,5- dihydropyrazin-2-yl]benzonitrile 30

4-[6-(4-aminopiperidin-1-yl)-3-(4- methoxyphenyl)-4-methyl-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 31

4-[3-(4-methoxyphenyl)-4-methyl- 5-oxo-6-{[(3S)-pyrrolidin-3-ylmethyl]amino}-4,5- dihydropyrazin-2-yl]benzonitrile 32

4-[3-(4-methoxyphenyl)-4-methyl- 5-oxo-6-{[(3R)-pyrrolidin-3-ylmethyl]amino}-4,5- dihydropyrazin-2-yl]benzonitrile 33

4-[6-(4-aminopiperidin-1-yl)-3-(4- cyclopropylphenyl)-4-methyl-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 34

4-[3-(4-cyclopropylphenyl)-4- methyl-5-oxo-6-{[(3S)-pyrrolidin-3-ylmethyl]amino}-4,5- dihydropyrazin-2-yl]benzonitrile 35

4-[3-(4-cyclopropylphenyl)-4- methyl-5-oxo-6-{[(3R)-pyrrolidin-3-ylmethyl]amino}-4,5- dihydropyrazin-2-yl]benzonitrile 36

4-[6-(4-amino-piperidin-1-yl)-4- (2-methoxy-ethyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]- benzonitrile 37

4-[6-(4-amino-piperidin-1-yl)-4- (2-hydroxy-ethyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]- benzonitrile 38

4-[6-(4-amino-piperidin-1-yl)-3- (4-cyclopropyl-phenyl)-4-(2-hydroxy-ethyl)-5-oxo-4,5-dihydro- pyrazin-2-yl]-benzonitrile 39

4-[6-(4-amino-piperidin-1-yl)-4- (3-methoxy-propyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]- benzonitrile 40

4-[6-(4-amino-piperidin-1-yl)-4- (3-hydroxy-propyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]- benzonitrile 41

4-[6-(4-amino-piperidin-1-yl)-4- (2-methoxy-ethyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]-2- fluoro-benzonitrile 42

4-[6-(4-amino-piperidin-1-yl)-4- (2-hydroxy-ethyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]-2- fluoro-benzonitrile 43

4-(6-{[((3S)-pyrrolidin-3- yl)methyl]amino}-4-(2- methoxyethyl)-3-(4-methylphenyl)-5-oxo(4- hydropyrazin-2-yl))-2- fluorobenzenecarbonitrile44

4-(6-{[((3S)-pyrrolidin-3- yl)methyl]amino}-4-(2- hydroxyethyl)-3-(4-methylphenyl)-5-oxo(4- hydropyrazin-2-yl))-2- fluorobenzenecarbonitrile45

4-[6-(4-aminopiperidyl)-4-(3- methoxypropyl)-3-(4-methylphenyl)-5-oxo(4- hydropyrazin-2-yl)]-2- fluorobenzenecarbonitrile46

4-[6-(4-aminopiperidyl)-4-(3- hydroxypropyl)-3-(4-methylphenyl)-5-oxo(4- hydropyrazin-2-yl)]-2- fluorobenzenecarbonitrile47

4-[6-(4-amino-piperidin-1-yl)-4- methyl-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]-2-fluoro- benzonitrile 48

4-[6-(4-amino-piperidin-1-yl)-4- methyl-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]-3-fluoro- benzonitrile 49

4-[6-(4-amino-piperidin-1-yl)-4- methyl-3-(6-methyl-pyridin-3-yl)-5-oxo-4,5-dihydro-pyrazin-2-yl]- benzonitrile 50

4-[6-(4-amino-piperidin-1-yl)-3- (4-cyclopropyl-phenyl)-4-methyl-5-oxo-4,5-dihydro-pyrazin-2-yl]- 2-fluoro-benzonitrile 51

5-[6-(4-amino-piperidin-1-yl)-3- (4-methoxy-phenyl)-4-methyl-5-oxo-4,5-dihydro-pyrazin-2-yl]- pyridine-2-carbonitrile 52

4-[6-(4-amino-piperidin-1-yl)-3- (4-methoxy-phenyl)-4-methyl-5-oxo-4,5-dihydro-pyrazin-2-yl]-2- fluoro-benzonitrile 53

5-[6-(4-amino-piperidin-1-yl)-3- (4-cyclopropyl-phenyl)-4-methyl-5-oxo-4,5-dihydro-pyrazin-2-yl]- pyridine-2-carbonitrile 54

5-{3-(4-methoxy-phenyl)-4- methyl-5-oxo-6-[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro- pyrazin-2-yl}-pyridine-2- carbonitrile55

2-fluoro-4-{3-(4-methoxy- phenyl)-4-methyl-5-oxo-6-[((3S)-pyrrolidin-3-ylmethyl)-amino]- 4,5-dihydro-pyrazin-2-yl}- benzonitrile56

5-{3-(4-ethyl-phenyl)-4-methyl-5- oxo-6-[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro- pyrazin-2-yl}-pyridine-2- carbonitrile 57

4-{3-(4-ethyl-phenyl)-4-methyl-5- oxo-6-[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro- pyrazin-2-yl}-2-fluoro- benzonitrile 58

5-{3-(4-cyclopropyl-phenyl)-4- methyl-5-oxo-6-[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro- pyrazin-2-yl}-pyridine-2- carbonitrile59

4-{3-(4-cyclopropyl-phenyl)-4- methyl-5-oxo-6-[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro- pyrazin-2-yl}-2-fluoro- benzonitrile 60

4-[6-(4-amino-piperidin-1-yl)-4- methyl-3-(1-methyl-1H-benzoimidazol-5-yl)-5-oxo-4,5- dihydro-pyrazin-2-yl]-2-fluoro-benzonitrile 61

4-{3-(4-cyclopropylmethoxy- phenyl)-4-methyl-5-oxo-6-[((3S)-pyrrolidin-3-ylmethyl)-amino]- 4,5-dihydro-pyrazin-2-yl}-2-fluoro-benzonitrile 62

4-{3-(4-ethoxy-phenyl)-4-methyl- 5-oxo-6-[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro- pyrazin-2-yl}-2-fluoro- benzonitrile 63

2-fluoro-4-[4-methyl-5-oxo-6- [((3S)-pyrrolidin-3-ylmethyl)-amino]-3-(4-trifluoromethoxy- phenyl)-4,5-dihydro-pyrazin-2-yl]-benzonitrile 64

2-Fluoro-4-[4-(3-hydroxy-propyl)- 6-(4-methylamino-piperidin-1-yl)-5-oxo-3-p-tolyl-4,5-dihydro- pyrazin-2-yl]-benzonitrile 65

2-Fluoro-4-[4-(2-hydroxy-ethyl)- 6-(4-methylamino-piperidin-1-yl)-5-oxo-3-p-tolyl-4,5-dihydro- pyrazin-2-yl]-benzonitrile 66

4-[6-(4-Amino-piperidin-1-yl)-4- methyl-3-(6-methyl-pyridin-3-yl)-5-oxo-4,5-dihydro-pyrazin-2-yl]- 2-fluoro-benzonitrile 67

4-(5-(4-aminopiperidin-1-yl)-2-(4- methoxyphenyl)-1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2- fluorobenzonitrile 68

5-(5-(4-aminopiperidin-1-yl)-2-(4- methoxyphenyl)-1-methyl-6-oxo-1,6-dihydropyridin-3- yl)picolinonitrile 69

(S)-5-(2-(4-methoxyphenyl)-1- methyl-6-oxo-5-(pyrrolidin-3-ylmethylamino)-1,6- dihydropyridin-3-yl)picolinonitrile 70

(S)-2-fluoro-4-(2-(4- methoxyphenyl)-1-methyl-6-oxo-5-(pyrrolidin-3-ylmethylamino)- 1,6-dihydropyridin-3- yl)benzonitrile 71

(S)-5-(2-(4-ethylphenyl)-1- methyl-6-oxo-5-(pyrrolidin-3-ylmethylamino)-1,6- dihydropyridin-3-yl)picolinonitrile 72

(S)-4-(2-(4-ethylphenyl)-1- methyl-6-oxo-5-(pyrrolidin-3-ylmethylamino)-1,6- dihydropyridin-3-yl)-2- fluorobenzonitrile 73

(S)-5-(2-(4-cyclopropylphenyl)-1- methyl-6-oxo-5-(pyrrolidin-3-ylmethylamino)-1,6- dihydropyridin-3-yl)picolinonitrile 74

(S)-4-(2-(4-cyclopropylphenyl)-1- methyl-6-oxo-5-(pyrrolidin-3-ylmethylamino)-1,6- dihydropyridin-3-yl)-2- fluorobenzonitrile 75

4-[5-(4-aminopiperidin-1-yl)-1- methyl-2-(4-methylphenyl)-6-oxopyridin-3-yl]benzonitrile 76

4-[5-(4-aminopiperidin-1-yl)-1- methyl-2-(4-methylphenyl)-6-oxopyridin-3-yl]-2- fluorobenzonitrile 77

4-[5-(4-aminopiperidin-1-yl)-2-(4- methoxyphenyl)-1-methyl-6-oxopyridin-3-yl]benzonitrile 78

4-{2-(4-methoxyphenyl)-1- methyl-5-[4- (methylamino)piperidyl]-6-oxo-3-hydropyridyl}benzenecarbonitrile 79

2-fluoro-4-{2-(4-methoxyphenyl)- 1-methyl-5-[4-(methylamino)piperidyl]-6-oxo(3- hydropyridyl)}benzenecarbonitrile 80

4-{1-methyl-5-[4- (methylamino)piperidyl]-2-(4- methylphenyl)-6-oxo-3-hydropyridyl}benzenecarbonitrile 81

4-[5-(4-aminopiperidin-1-yl)-1- (cyclopropylmethyl)-2-(4-methoxyphenyl)-6-oxopyridin-3- yl]benzonitrile 82

4-[5-(4-aminopiperidin-1-yl)-1- ethyl-2-(4-methoxyphenyl)-6-oxopyridin-3-yl]benzonitrile 83

4-[5-(4-aminopiperidin-1-yl)-2-(3- fluoro-4-methoxyphenyl)-1-methyl-6-oxopyridin-3- yl]benzonitrile 84

4-[5-(4-aminopiperidin-1-yl)-2-(3- fluoro-4-methoxyphenyl)-1-methyl-6-oxopyridin-3-yl]-2- fluorobenzonitrile 85

4-[5-(4-aminopiperidin-1-yl)-1- (cyclopropylmethyl)-2-(4-methoxyphenyl)-6-oxopyridin-3- yl]-2-fluorobenzonitrile 86

2-fluoro-4-[2-(3-fluoro-4- methoxyphenyl)-1-methyl-5-[4-(methylamino)piperidin-1-yl]-6- oxopyridin-3-yl]benzonitrile 87

4-[5-(4-aminopiperidin-1-yl)-1- ethyl-2-(4-methoxyphenyl)-6-oxopyridin-3-yl]-2- fluorobenzonitrile 88

2-[3-(4-aminopiperidyl)-5-(4- cyano-3-fluorophenyl)-6-(4-methoxyphenyl)-2- oxohydropyrazinyl]acetamide 89

3-[3-(4-aminopiperidyl)-5-(4- cyano-3-fluorophenyl)-6-(4-methoxyphenyl)-2- oxohydropyrazinyl]propanamide 90

4-[3-(4-aminopiperidyl)-5-(4- cyano-3-fluorophenyl)-6-(4-methoxyphenyl)-2- oxohydropyrazinyl]butanamide 91

4-{5-(4-aminopiperidyl)-2-[6- (dimethylamino)(3-pyridyl)]-1-methyl-6-oxo(3-hydropyridyl)}-2- fluorobenzenecarbonitrile 92

4-{2-[6-(dimethylamino)(3- pyridyl)]-1-methyl-5-[4-(methylamino)piperidyl]-6-oxo-3- hydropyridyl}benzenecarbonitrile

In some embodiments, the substituted heterocyclic derivative compounddescribed herein has the structure provided in Table 2.

TABLE 2

Preparation of the Substituted Heterocyclic Derivative Compounds

The compounds used in the reactions described herein are made accordingto organic synthesis techniques known to those skilled in this art,starting from commercially available chemicals and/or from compoundsdescribed in the chemical literature. “Commercially available chemicals”are obtained from standard commercial sources including Acros Organics(Pittsburgh, Pa.), Aldrich Chemical (Milwaukee, Wis., including SigmaChemical and Fluka), Apin Chemicals Ltd. (Milton Park, UK), AvocadoResearch (Lancashire, U.K.), BDH Inc. (Toronto, Canada), Bionet(Cornwall, U.K.), Chemservice Inc. (West Chester, Pa.), CrescentChemical Co. (Hauppauge, N.Y.), Eastman Organic Chemicals, Eastman KodakCompany (Rochester, N.Y.), Fisher Scientific Co. (Pittsburgh, Pa.),Fisons Chemicals (Leicestershire, UK), Frontier Scientific (Logan,Utah), ICN Biomedicals, Inc. (Costa Mesa, Calif.), Key Organics(Cornwall, U.K.), Lancaster Synthesis (Windham, N.H.), MaybridgeChemical Co. Ltd. (Cornwall, U.K.), Parish Chemical Co. (Orem, Utah),Pfaltz & Bauer, Inc. (Waterbury, Conn.), Polyorganix (Houston, Tex.),Pierce Chemical Co. (Rockford, Ill.), Riedel de Haen AG (Hanover,Germany), Spectrum Quality Product, Inc. (New Brunswick, N.J.), TCIAmerica (Portland, Oreg.), Trans World Chemicals, Inc. (Rockville, Md.),and Wako Chemicals USA, Inc. (Richmond, Va.).

Methods known to one of ordinary skill in the art are identified throughvarious reference books and databases. Suitable reference books andtreatise that detail the synthesis of reactants useful in thepreparation of compounds described herein, or provide references toarticles that describe the preparation, include for example, “SyntheticOrganic Chemistry”, John Wiley & Sons, Inc., New York; S. R. Sandler etal., “Organic Functional Group Preparations,” 2nd Ed., Academic Press,New York, 1983; H. O. House, “Modern Synthetic Reactions”, 2nd Ed., W.A. Benjamin, Inc. Menlo Park, Calif. 1972; T. L. Gilchrist,“Heterocyclic Chemistry”, 2nd Ed., John Wiley & Sons, New York, 1992; J.March, “Advanced Organic Chemistry: Reactions, Mechanisms andStructure”, 4th Ed., Wiley-Interscience, New York, 1992. Additionalsuitable reference books and treatise that detail the synthesis ofreactants useful in the preparation of compounds described herein, orprovide references to articles that describe the preparation, includefor example, Fuhrhop, J. and Penzlin G. “Organic Synthesis: Concepts,Methods, Starting Materials”, Second, Revised and Enlarged Edition(1994) John Wiley & Sons ISBN: 3-527-29074-5; Hoffman, R. V. “OrganicChemistry, An Intermediate Text” (1996) Oxford University Press, ISBN0-19-509618-5; Larock, R. C. “Comprehensive Organic Transformations: AGuide to Functional Group Preparations” 2nd Edition (1999) Wiley-VCH,ISBN: 0-471-19031-4; March, J. “Advanced Organic Chemistry: Reactions,Mechanisms, and Structure” 4th Edition (1992) John Wiley & Sons, ISBN:0-471-60180-2; Otera, J. (editor) “Modern Carbonyl Chemistry” (2000)Wiley-VCH, ISBN: 3-527-29871-1; Patai, S. “Patai's 1992 Guide to theChemistry of Functional Groups” (1992) Interscience ISBN: 0-471-93022-9;Solomons, T. W. G. “Organic Chemistry” 7th Edition (2000) John Wiley &Sons, ISBN: 0-471-19095-0; Stowell, J. C., “Intermediate OrganicChemistry” 2nd Edition (1993) Wiley-Interscience, ISBN: 0-471-57456-2;“Industrial Organic Chemicals: Starting Materials and Intermediates: AnUllmann's Encyclopedia” (1999) John Wiley & Sons, ISBN: 3-527-29645-X,in 8 volumes; “Organic Reactions” (1942-2000) John Wiley & Sons, in over55 volumes; and “Chemistry of Functional Groups” John Wiley & Sons, in73 volumes.

Specific and analogous reactants may also be identified through theindices of known chemicals prepared by the Chemical Abstract Service ofthe American Chemical Society, which are available in most public anduniversity libraries, as well as through on-line databases (the AmericanChemical Society, Washington, D.C., may be contacted for more details).Chemicals that are known but not commercially available in catalogs maybe prepared by custom chemical synthesis houses, where many of thestandard chemical supply houses (e.g., those listed above) providecustom synthesis services. A reference for the preparation and selectionof pharmaceutical salts of the substituted heterocyclic derivativecompounds described herein is P. H. Stahl & C. G. Wermuth “Handbook ofPharmaceutical Salts”, Verlag Helvetica Chimica Acta, Zurich, 2002.

The substituted heterocyclic derivative compounds are prepared by thegeneral synthetic routes described below in Schemes 1-3.

The N-substituted pyrazinone derivative compounds are prepared by thegeneral synthetic route described below in Scheme 1.

Referring to Scheme 1, displacement of chloroacetonitrile is carried outwith a variety of amines J-NH₂ under basic conditions to form compoundK. Compound L is obtained from treating compound K to Vielsmeircondition. Selective displacement of the dichloride compound L iscarried out with a variety of amines MM′-NH under basic conditions toform compound N. Compound R is prepared from aryl halide compound Nusing palladium-mediated cross coupling conditions with boronic acidsQ-(OH)₂.

The substituted pyrazinone derivative compounds are prepared by thegeneral synthetic route described below in Scheme 2.

Referring to Scheme 2, compound V is obtained from condensation of avariety of aldehydes T-CHO and amines U—NH₂, followed by cyanideaddition. Compound X is obtained from compound V under Vielsmeircondition. Selective displacement of dichloride compound X is carriedout with a variety of amines YY′—NH under basic conditions to formcompound Z. Compound AB is prepared from aryl halide compound Z usingpalladium-mediated cross coupling conditions with boronic acidsAA-B(OH)₂.

The substituted pyridone derivative compounds are prepared by thegeneral synthetic route described below in Scheme 3.

Referring to Scheme 3, compound AO is obtained from N-alkylation ofcompound AM with a variety of alkyl halides AN—X. Compound AQ isprepared from aryl halide compound AO using aryl coupling conditions,such as Suzuki conditions with boronic acids AP—B(OH)₂. Bromination ofcompound AQ afforded compound AR. Selective displacement of dibromidecompound AR is carried out with a variety of amines ASAS′—NH under basicconditions, or via Buchwald coupling condition, to form compound AT.Compound AV is prepared from aryl halide compound AT usingpalladium-mediated cross coupling conditions with boronic acidsAU—B(OH)₂.

Pharmaceutical Compositions

In certain embodiments, the substituted heterocyclic derivative compoundas described herein is administered as a pure chemical. In otherembodiments, the substituted heterocyclic derivative compound describedherein is combined with a pharmaceutically suitable or acceptablecarrier (also referred to herein as a pharmaceutically suitable (oracceptable) excipient, physiologically suitable (or acceptable)excipient, or physiologically suitable (or acceptable) carrier) selectedon the basis of a chosen route of administration and standardpharmaceutical practice as described, for example, in Remington: TheScience and Practice of Pharmacy (Gennaro, 21^(st) Ed. Mack Pub. Co.,Easton, Pa. (2005)), the disclosure of which is hereby incorporatedherein by reference in its entirety.

Accordingly, provided herein is a pharmaceutical composition comprisingat least one substituted heterocyclic derivative compound as describedherein, or a stereoisomer, pharmaceutically acceptable salt, hydrate,solvate, or N-oxide thereof, together with one or more pharmaceuticallyacceptable carriers. The carrier(s) (or excipient(s)) is acceptable orsuitable if the carrier is compatible with the other ingredients of thecomposition and not deleterious to the recipient (i.e., the subject) ofthe composition.

One embodiment provides a pharmaceutical composition comprising acompound of Formula (I), or a pharmaceutically acceptable salt thereof,and a pharmaceutically acceptable excipient.

In certain embodiments, the substituted heterocyclic derivative compoundas described by Formula (I) is substantially pure, in that it containsless than about 5%, or less than about 1%, or less than about 0.1%, ofother organic small molecules, such as contaminating intermediates orby-products that are created, for example, in one or more of the stepsof a synthesis method.

Suitable oral dosage forms include, for example, tablets, pills,sachets, or capsules of hard or soft gelatin, methylcellulose or ofanother suitable material easily dissolved in the digestive tract.Suitable nontoxic solid carriers can be used which include, for example,pharmaceutical grades of mannitol, lactose, starch, magnesium stearate,sodium saccharin, talcum, cellulose, glucose, sucrose, magnesiumcarbonate, and the like. (See, e.g., Remington: The Science and Practiceof Pharmacy (Gennaro, 21^(st) Ed. Mack Pub. Co., Easton, Pa. (2005)).

The dose of the composition comprising at least one substitutedheterocyclic derivative compound as described herein may differ,depending upon the patient's (e.g., human) condition, that is, stage ofthe disease, general health status, age, and other factors that a personskilled in the medical art will use to determine dose.

Pharmaceutical compositions may be administered in a manner appropriateto the disease to be treated (or prevented) as determined by personsskilled in the medical arts. An appropriate dose and a suitable durationand frequency of administration will be determined by such factors asthe condition of the patient, the type and severity of the patient'sdisease, the particular form of the active ingredient, and the method ofadministration. In general, an appropriate dose and treatment regimenprovides the composition(s) in an amount sufficient to providetherapeutic and/or prophylactic benefit (e.g., an improved clinicaloutcome, such as more frequent complete or partial remissions, or longerdisease-free and/or overall survival, or a lessening of symptomseverity. Optimal doses may generally be determined using experimentalmodels and/or clinical trials. The optimal dose may depend upon the bodymass, weight, or blood volume of the patient.

Oral doses can typically range from about 1.0 mg to about 1000 mg, oneto four times, or more, per day.

Biology

Epigenetics is the study of heritable changes in gene expression causedby mechanisms other than the underlying DNA sequence. Molecularmechanisms that play a role in epigenetic regulation include DNAmethylation and chromatin/histone modifications.

The genomes of eukaryotic organisms are highly organized within thenucleus of the cell. Tremendous compaction is required to package the 3billion nucleotides of the human genome into the nucleus of a cell.Chromatin is the complex of DNA and protein that makes up chromosomes.Histones are the major protein component of chromatin, acting as spoolsaround which DNA winds. Changes in chromatin structure are affected bycovalent modifications of histone proteins and by non-histone bindingproteins. Several classes of enzymes are known which can modify histonesat various sites.

There are a total of six classes of histones (HI, H2A, H2B, H3, H4, andH5) organized into two groups: core histones (H2A, H2B, H3, and H4) andlinker histones (HI and H5). The basic unit of chromatin is thenucleosome, which consists of about 147 base pairs of DNA wrapped aroundthe core histone octamer, consisting of two copies each of the corehistones H2A, H2B, H3, and H4.

Basic nucleosome units are then further organized and condensed by theaggregation and folding of nucleosomes to form a highly condensedchromatin structure. A range of different states of condensation arepossible, and the tightness of chromatin structure varies during thecell cycle, being most compact during the process of cell division.

Chromatin structure plays a critical role in regulating genetranscription, which cannot occur efficiently from highly condensedchromatin. The chromatin structure is controlled by a series of posttranslational modifications to histone proteins, notably histones H3 andH4, and most commonly within the histone tails which extend beyond thecore nucleosome structure. These modifications acetylation, methylation,phosphorylation, ribosylation sumoylation, ubiquitination,citrullination, deimination, and biotinylation. The core of histones H2Aand H3 can also be modified. Histone modifications are integral todiverse biological processes such as gene regulation, DNA repair, andchromosome condensation.

Histone methylation is one of the most important chromatin marks; theseplay important roles in transcriptional regulation, DNA-damage response,heterochromatin formation and maintenance, and X-chromosomeinactivation. A recent discovery also revealed that histone methylationaffects the splicing outcome of pre-mRNA by influencing the recruitmentof splicing regulators. Histone methylation includes mono-, di-, andtri-methylation of lysines, and mono-, symmetric di-, and asymmetricdi-methylation of arginines. These modifications can be either anactivating or repressing mark, depending on the site and degree ofmethylation.

Histone Demethylases

A “demethylase” or “protein demethylase,” as referred to herein, refersto an enzyme that removes at least one methyl group from polypeptide.Demethylases comprise a JmjC domain, and can be a methyl-lysine ormethyl-arginine demethylase. Some demethylases act on histones, e.g.,act as a histone H3 or H4 demethylase. For example, an H3 demethylasemay demethylate one or more of H3K4, H3K9, H3K27, H3K36 and/or H3K79.Alternately, an H4 demethylase may demethylate histone H4K20.Demethylases are known which can demethylate either a mono-, di- and/ora tri-methylated substrate. Further, histone demethylases can act on amethylated core histone substrate, a mononucleosome substrate, adinucleosome substrate and/or an oligonucleosome substrate, peptidesubstrate and/or chromatin (e.g., in a cell-based assay).

The first lysine demethylase discovered was lysine specific demethylase1 (LSD1/KDM1), which demethylates both mono- and di-methylated H3K4 orH3K9, using flavin as a cofactor. A second class of Jumonji C (JmjC)domain containing histone demthylases were predicted, and confirmed whena H3K36 demethylase was found used a formaldehyde release assay, whichwas named JmjC domain containing histone demethylase 1 (JHDM1/KDM2A).

More JmjC domain-containing proteins were subsequently identified andthey can be phylogenetically clustered into seven subfamilies: JHDM1,JHDM2, JHDM3, JMJD2, JARID, PHF2/PHF8, UTX/UTY, and JmjC domain only.

LSD-1

Lysine-specific demethylase 1 (LSD1) is a histone lysine demethylasethat specifically demethylates monomethylated and dimethylated histoneH3 at K4 and also demethylates dimethylated histone H3 at K9. Althoughthe main target of LSD1 appears to be mono- and di-methylated histonelysines, specifically H3K4 and H3K9, there is evidence in the literaturethat LSD 1 can demethylate methylated lysines on non-histone proteinslike p53, E2F1, Dnmtl and STAT3.

LSD 1 has a fair degree of structural similarity and amino acididentity/homology to polyamine oxidases and monoamine oxidases, all ofwhich (i.e., MAO-A, MAO-B and LSD1) are flavin dependent amine oxidaseswhich catalyze the oxidation of nitrogen-hydrogen bonds and/ornitrogen-carbon bonds. LSD1 also includes an N-terminal SWRIM domain.There are two transcript variants of LSD1 produced by alternativesplicing.

Methods of Use

In some embodiments, the compounds disclosed herein are capable ofinhibiting LSD1 activity in a biological sample by contacting thebiological sample with a substituted heterocyclic compound as disclosedherein. In some embodiments, a substituted heterocyclic compound asdisclosed herein is capable of modulating the level of histone 4 lysine3 methylation in the biological sample. In some embodiments, asubstituted heterocyclic compound as disclosed herein is capable ofmodulating histone-3 lysine-9 methylation levels in the biologicalsample.

The substituted heterocyclic compounds disclosed herein lack significantMAO-A or MAO-B inhibitory activity. In some embodiments, the substitutedheterocyclic compound as disclosed herein inhibits LSD1 inhibitoryactivity to a greater extent than MAO-A and/or MAO-B inhibitoryactivity.

One embodiment provides a method of regulating gene transcription in acell comprising inhibiting lysine-specific demethylase 1 activity byexposing the lysine-specific demethylase 1 enzyme to a compound ofFormula (I).

Methods of Treatment

Disclosed herein are methods of modulating demethylation in a cell or ina subject, either generally or with respect to one or more specifictarget genes. Demethylation can be modulated to control a variety ofcellular functions, including without limitation: differentiation;proliferation; apoptosis; tumorigenesis, leukemogenesis or otheroncogenic transformation events; hair loss; or sexual differentiation.

One embodiment provides a method of treating cancer in a patient in needthereof, comprising administering to the patient a compound of Formula(I), or a pharmaceutically acceptable salt thereof.

In a further embodiment is the method for treating cancer in a subjectwherein the cancer is selected from prostate cancer, breast cancer,bladder cancer, lung cancer or melanoma.

Other embodiments and uses will be apparent to one skilled in the art inlight of the present disclosures. The following examples are providedmerely as illustrative of various embodiments and shall not be construedto limit the invention in any way.

EXAMPLES I. Chemical Synthesis

Unless otherwise noted, reagents and solvents were used as received fromcommercial suppliers. Anhydrous solvents and oven-dried glassware wereused for synthetic transformations sensitive to moisture and/or oxygen.Yields were not optimized. Reaction times are approximate and were notoptimized. Column chromatography and thin layer chromatography (TLC)were performed on silica gel unless otherwise noted. Spectra are givenin ppm (δ) and coupling constants, J are reported in Hertz. For protonspectra the solvent peak was used as the reference peak.

Preparation 1A: benzylamino-acetonitrile

A mixture of benzylamine (1.72 g, 16 mmol), chloroacetonitrile (1.81 g,24 mmol) and K₂CO₃ (4.4 g, 32 mmol) in acetonitrile (50 mL) was heatedat 60° C. overnight. The reaction mixture was filtered and the filtratewas concentrated. The residue was purified by silica chromatography(5:1, PE:EA) to give 1.8 g (77%) of the title compound as the HCl salt.[M+H] Calc'd for C₉H₁₀N₂, 147; Found, 147.

Preparation 1B: 1-benzyl-3,5-dichloropyrazin-2(1H)-one

To a mixture of 2-(benzylamino)acetonitrile, HCl (1 g, 5.5 mmol) in 1,2-dichlorobenzene (15 mL) was added (COCl)₂ (3.47 g, 27.4 mmol). Themixture was stirred at 90° C. for 1 h. The reaction mixture was cooledto 40° C., and 4 drops DMF were added. The mixture was stirred heated at125° C. for 3 h. The solvent was removed in vacuo, and the residue waspurified by silica chromatography (1:5, EA:PE) to give 1.0 g (71.4%) ofthe title compound. ¹H NMR (400 MHz, CDCl₃): δ 5.11 (s, 2H), 7.18 (s,1H), 7.33-7.43 (m, 5H).

Preparation 1C: tert-butyl1-(4-benzyl-6-chloro-3-oxo-3,4-dihydropyrazin-2-yl)piperidin-4-ylcarbamate

To a solution of 1-benzyl-3,5-dichloropyrazin-2(1H)-one (500 mg, 1.96mmol) in DMF (5 mL) was added (tert-butoxy)-N-(4-piperidyl)carboxamide(392 mg, 1.96 mmol) and DIEA (506 mg, 3.92 mmol). The mixture wasstirred at 120° C. for 2 h. DMF was concentrated in vacuo and theresidue partitioned between water and EA. The organic phase was washedwith brine, dried and concentrated to give 550 mg (66.9%) of the titleproduct. ¹H NMR (400 MHz, CDCl₃): δ 1.42-1.51 (m, 11H), 2.02-2.05 (m,2H), 3.00-3.06 (m, 2H), 3.70 (d, J=2.0 Hz, 1H), 4.47 (d, J=5.6 Hz, 1H),4.76 (d, J=12.8 Hz, 2H), 4.98 (s, 2H), 6.66 (s, 1H), 7.29-7.79 (m, 5H).

Preparation 1D: tert-butyl1-(4-benzyl-6-(4-cyanophenyl)-3-oxo-3,4-dihydropyrazin-2-yl)piperidin-4-ylcarbamate

To a solution of tert-butyl1-(4-benzyl-6-chloro-3-oxo-3,4-dihydropyrazin-2-yl)piperidin-4-ylcarbamate(360 mg, 0.86 mmol) in DMF (5 mL) was added 4-cyanophenylboronic acid(253 mg, 1.72 mmol),[1,1′-Bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (56 mg,0.086 mmol) and K₂CO₃ (237 mg, 1.72 mmol) under N₂ atmosphere. Themixture was stirred at 145° C. for 4 h. Water was added and the productextracted with EtOAc (3×). The organics were combined, washed withbrine, dried and concentrated. The residue was purified by columnchromatography (1:5, EA:PE) to give 300 mg (71.8%) of the titlecompound. ¹H NMR (400 MHz, CDCl₃): δ 1.46-1.55 (m, 11H), 2.04-2.09 (m,2H), 3.07-3.12 (m, 2H), 3.73-3.75 (m, 1H), 4.47-4.49 (m, 1H), 4.76 (d,J=13.2 Hz, 2H), 5.12 (s, 2H), 7.21 (s, 1H), 7.33-7.37 (m, 5H), 7.63 (d,J=8.4 Hz, 2H), 7.79 (d, J=8.4 Hz, 2H).

Example 1:4-[6-(4-aminopiperidin-1-yl)-4-benzyl-5-oxo-4,5-dihydropyrazin-2-yl]benzonitrile

To a solution of tert-butyl1-(4-benzyl-6-(4-cyanophenyl)-3-oxo-3,4-dihydropyrazin-2-yl)piperidin-4-ylcarbamate(300 mg, 0.62 mmol) in EtOAc (5 mL) was added HCl/EA (10 mL), themixture was stirred at RT for 2 h. The solvent was concentrated in vacuoand the residue was purified by preparative HPLC to give 169 mg (71%) ofthe title compound as the HCl salt. ¹H-NMR (CD₃OD, 400 MHz): δ 1.76-1.83(m, 2H), 2.12-2.15 (m, 2H), 3.04-3.11 (t, J=12.6 Hz, 2H) 3.44-3.46 (m,1H), 3.91-3.96 (m, 2H), 5.21 (s, 2H), 7.33-7.44 (m, 5H), 7.75 (d, J=8.4Hz, 2H), 7.89 (s, 1H), 8.01 (d, J=8.4 Hz, 2H). [M+H] Calc'd forC₂₃H₂₃N₅O, 386; Found, 386.

Example 2:4-[6-(4-aminopiperidin-1-yl)-4-[(4-methylphenyl)methyl]-5-oxo-4,5-dihydropyrazin-2-yl]benzonitrile

The title compound was prepared as the HCl salt in 18% overall yieldaccording to the general procedure for the preparation of Example 1. ¹HNMR (400 MHz, CD₃OD): δ 1.68-1.72 (m, 2H), 2.04-2.07 (m, 2H), 2.27 (s,3H), 2.99 (t, J=12.4 Hz, 2H), 3.26-3.27 (m, 1H), 4.84-4.87 (m, 2H), 5.09(s, 2H), 7.12 (d, J=8.0 Hz, 2H), 7.24 (d, J=8.0 Hz, 2H), 7.67 (d, J=8.8Hz, 2H), 7.79 (s, 1H), 7.94 (d, J=8.4 Hz, 2H). [M+H] Calc'd forC₂₄H₂₅N₅O, 400; Found, 400.

Example 3:4-[6-(4-aminopiperidin-1-yl)-4-[(2-fluorophenyl)methyl]-5-oxo-4,5-dihydropyrazin-2-yl]benzonitrile

The title compound was prepared as the HCl salt in 14% overall yieldaccording to the general procedure for the preparation of Example 1. ¹HNMR (400 MHz, CD₃OD): δ 1.67-1.74 (m, 2H), 2.03-2.06 (m, 2H), 2.97 (t,J=12.2 Hz, 2H), 3.33-3.39 (m, 1H), 4.84-4.87 (m, 2H), 5.20 (s, 2H),7.09-7.15 (m, 2H), 7.30-7.35 (m, 2H), 7.69 (d, J=8.0 Hz, 2H), 7.80 (s,1H), 7.96 (d, J=8.4 Hz, 2H). [M+H] Calc'd for C₂₃H₂₂FN₅O, 404; Found,404.

Example 4:4-[6-(4-aminopiperidin-1-yl)-4-(3-chloro-benzyl)-5-oxo-4,5-dihydro-pyrazin-2-yl]-benzonitrile

The title compound was prepared as the HCl salt in 6% overall yieldaccording to the general procedure for the preparation of Example 1. ¹HNMR (400 MHz, CD₃OD): δ 1.70-1.80 (m, 2H), 2.08-2.12 (m, 2H), 3.03 (t,J=12.0 Hz, 2H), 3.39-3.44 (m, 1H), 4.89-4.93 (m, 2H), 5.16 (s, 2H),7.30-7.35 (m, 3H), 7.42 (s, 1H), 7.72 (d, J=8.4 Hz, 2H), 7.89 (s, 1H),8.01 (d, J=8.4 Hz, 2H). [M+H] Calc'd for C₂₃H₂₂ClN₅O, 420; Found, 420.

Example 5:4-[6-(4-amino-piperidin-1-yl)-4-methyl-5-oxo-4,5-dihydro-pyrazin-2-yl]-2-fluoro-benzonitrile

The title compound was prepared as the HCl salt in 17% overall yieldaccording to the general procedure for the preparation of Example 1. ¹HNMR (300 MHz, DMSO-d₆): δ 1.57-1.66 (m, 2H), 1.98-2.03 (m, 2H),2.93-2.97 (m, 2H), 3.27-3.30 (m, 1H), 3.47 (s, 3H), 4.73-4.78 (m, 2H),7.85-7.97 (m, 3H), 8.10-8.18 (m, 4H). [M+H] Calc'd for C₁₇H₁₈FN₅O, 328;Found, 328.

Example 6:4-[6-(4-Amino-piperidin-1-yl)-4-ethyl-5-oxo-4,5-dihydro-pyrazin-2-yl]-2-fluoro-benzonitrile

The title compound was prepared as the HCl salt in 15% overall yieldaccording to the general procedure for the preparation of Example 1. ¹HNMR (400 MHz, CD₃OD): δ 1.37 (t, J=7.2 Hz, 3H), 1.70-1.79 (m, 2H),2.09-2.11 (m, 2H), 2.98-3.05 (m, 2H), 3.38-3.43 (m, 1H), 4.00-4.05 (q,J=7.0 Hz, 2H), 4.86-4.89 (m, 2H), 7.70-7.74 (m, 1H), 7.82 (s, 1H),7.83-7.84 (m, 1H), 7.88 (s, 1H). [M+H] Calc'd for C₁₈H₂₀FN₅O, 342;Found, 342.

Example 7:4-[6-(4-amino-piperidin-1-yl)-4-cyclopropylmethyl-5-oxo-4,5-dihydro-pyrazin-2-yl]-2-fluoro-benzonitrile

The title compound was prepared as the HCl salt in 14% overall yieldaccording to the general procedure for the preparation of Example 1. ¹HNMR (400 MHz, DMSO-d₆): δ 0.43-0.53 (m, 4H), 1.28-1.32 (m, 1H),1.58-1.67 (m, 2H), 2.00-2.03 (m, 2H), 2.50-2.53 (m, 1H), 2.95-3.01 (m,2H), 3.76 (d, J=6.8 Hz, 2H), 4.76 (d, J=12.8 Hz, 2H), 7.89-7.98 (m, 3H),8.17-8.21 (m, 4H). [M+H] Calc'd for C₂₀H₂₂FN₅O, 368; Found, 368.

Example 8:4-[6-(4-aminopiperidin-1-yl)-4-[(3-fluorophenyl)methyl]-5-oxo-4,5-dihydropyrazin-2-yl]benzonitrile

The title compound was prepared as the HCl salt in 17% overall yieldaccording to the general procedure for the preparation of Example 1. ¹HNMR (400 MHz, DMSO-d₆): δ 1.60-1.69 (m, 2H), 1.98-2.01 (m, 2H),2.89-2.95 (m, 2H), 3.28-3.34 (m, 1H), 4.78-4.83 (m, 2H), 5.08 (s, 2H),6.92-7.12 (m, 3H), 7.24-7.30 (m, 1H), 7.62 (d, J=8.0 Hz, 2H), 7.79 (s,1H), 7.91 (d, J=8.0 Hz, 2H). [M+H] Calc'd for C₂₃H₂₃FN₅O, 404; Found,404.

Example 9:4-[6-(4-aminopiperidin-1-yl)-4-[(4-fluorophenyl)methyl]-5-oxo-4,5-dihydropyrazin-2-yl]benzonitrile

The title compound was prepared as the HCl salt in 15% overall yieldaccording to the general procedure for the preparation of Example 1. ¹HNMR (400 MHz, DMSO-d₆): δ 1.83-1.87 (m, 2H), 2.18-2.22 (m, 2H),3.11-3.17 (m, 2H), 3.41-3.48 (m, 1H), 4.96-5.04 (m, 2H), 5.28 (s, 2H),7.21 (t, J=8.8 Hz, 2H), 7.56-7.59 (m, 2H), 7.85 (d, J=8.0 Hz, 2H), 8.01(s, 1H), 8.13 (d, J=8.0 Hz, 2H). [M+H] Calc'd for C₂₃H₂₃FN₅O, 404;Found, 404.

Example 10:4-[6-(4-aminopiperidin-1-yl)-4-[(3-methoxyphenyl)methyl]-5-oxo-4,5-dihydropyrazin-2-yl]benzonitrile

The title compound was prepared as the HCl salt in 18% overall yieldaccording to the general procedure for the preparation of Example 1. ¹HNMR (400 MHz, CD₃OD): δ 1.72-1.82 (m, 2H), 2.11-2.14 (m, 2H), 3.06 (t,J=12.0 Hz, 2H), 3.41-3.47 (m, 1H), 3.80 (s, 3H), 4.91-4.94 (m, 2H), 5.18(s, 2H), 6.89-6.98 (m, 3H), 7.29 (t, J=8.4 Hz, 1H), 7.74 (d, J=8.4 Hz,2H), 7.88 (s, 1H), 8.02 (d, J=8.4 Hz, 2H). [M+H] Calc'd for C₂₄H₂₆N₅O₂,416; Found, 416.

Example 11:4-[6-(4-aminopiperidin-1-yl)-4-[(4-methoxyphenyl)methyl]-5-oxo-4,5-dihydropyrazin-2-yl]benzonitrile

The title compound was prepared as the HCl salt in 17% overall yieldaccording to the general procedure for the preparation of Example 1. ¹HNMR (400 MHz, CD₃OD): δ 1.58-1.68 (m, 2H), 1.93-1.99 (m, 2H), 2.99 (t,J=12.8 Hz, 2H), 3.26-3.32 (m, 1H), 3.67 (s, 3H), 4.78-4.83 (m, 2H), 5.01(s, 2H), 6.80 (d, J=8.4 Hz, 2H), 7.25 (d, J=8.4 Hz, 2H), 7.62 (d, J=8.4Hz, 2H), 7.76 (s, 1H), 7.90 (d, J=8.4 Hz, 2H). [M+H] Calc'd forC₂₄H₂₆N₅O₂, 416; Found, 416.

Example 12:4-[6-(4-aminopiperidin-1-yl)-5-oxo-4-[(1R)-1-phenylethyl]-4,5-dihydropyrazin-2-yl]benzonitrile

The title compound was prepared as the HCl salt in 14% overall yieldaccording to the general procedure for the preparation of Example 1. ¹HNMR (400 MHz, CD₃OD): δ 1.63-1.74 (m, 5H), 2.00-2.03 (m, 2H), 2.91-2.99(m, 2H), 3.03-3.36 (m, 1H), 4.78-4.84 (m, 2H), 6.14-6.19 (m, 1H),7.20-7.33 (m, 5H), 7.41 (s, 1H), 7.59 (d, J=8.4 Hz, 2H), 7.81 (d, J=8.4Hz, 2H). [M+H] Calc'd for C₂₄H₂₆N₅O, 400; Found, 400.

Example 13:4-[6-(4-aminopiperidin-1-yl)-5-oxo-4-[(1S)-1-phenylethyl]-4,5-dihydropyrazin-2-yl]benzonitrile

The title compound was prepared as the HCl salt in 16% overall yieldaccording to the general procedure for the preparation of Example 1. ¹HNMR (400 MHz, CD₃OD): δ 1.63-1.74 (m, 5H), 2.00-2.03 (m, 2H), 2.91-2.99(m, 2H), 3.03-3.36 (m, 1H), 4.78-4.84 (m, 2H), 6.14-6.19 (m, 1H),7.20-7.33 (m, 5H), 7.41 (s, 1H), 7.59 (d, J=8.4 Hz, 2H), 7.81 (d, J=8.4Hz, 2H). [M+H] Calc'd for C₂₄H₂₆N₅O, 400; Found, 400.

Example 14:4-[6-(4-Amino-piperidin-1-yl)-5-oxo-4-(tetrahydro-furan-3-ylmethyl)-4,5-dihydro-pyrazin-2-yl]-2-fluoro-benzonitrile

The title compound was prepared as the HCl salt in 66% overall yieldaccording to the general procedure for the preparation of Example 1. ¹HNMR (400 MHz, CD₃OD): δ 1.59-1.70 (m, 3H), 1.91-2.03 (m, 3H), 2.71-2.78(m, 1H), 2.93 (t, J=12.0 Hz, 2H), 3.29-3.35 (m, 1H), 3.50-3.54 (m, 1H),3.64-3.70 (m, 2H), 3.82-3.98 (m, 3H), 4.79-4.82 (m, 2H), 7.62-7.79 (m,4H). [M+H] Calc'd for C₂₁H₂₄FN₅O₂, 398; Found, 398.

Example 15:4-[6-(4-Amino-piperidin-1-yl)-5-oxo-4-(tetrahydro-pyran-4-ylmethyl)-4,5-dihydro-pyrazin-2-yl]-2-fluoro-benzonitrile

The title compound was prepared as the HCl salt in 47% overall yieldaccording to the general procedure for the preparation of Example 1. ¹HNMR (400 MHz, CD₃OD): δ 1.31-1.37 (m, 2H), 1.45-1.48 (m, 2H), 1.63-1.67(m, 2H), 2.00-2.07 (m, 3H), 2.93 (t, J=12.0 Hz, 2H), 3.25-3.33 (m, 3H),3.79-3.87 (m, 4H), 4.77-4.81 (m, 2H), 7.62-7.76 (m, 4H). [M+H] Calc'dfor C₂₂H₂₆FN₅O₂, 412; Found, 412.

Preparation 16A: 2-(methylamino)-2-(4-methylphenyl)ethanenitrile

To a solution of 4-methylbenzaldehyde (10.0 g, 83.3 mmol) in methanol(200 mL) was added methylamine (20.7 g, 25%, 166.6 mmol). The mixturewas stirred at RT for 1 h. TMS-CN (12.4 g, 125 mmol) was added and themixture was stirred overnight. Aq. NH₄Cl was added and the reactionmixture was extracted with EA (3×). The organics were combined, washedwith brine, dried and concentrated. The residue was taken in ether whereHCl gas was bubbled in for 5 min. The resulted precipitate was filtered,washed with ether and dried to give 13.5 g (82%) of the title compoundas the HCl salt. [M+H] Calc'd for C₁₀H₁₂N₂, 161; Found, 161.

Preparation 16B:3,5-dichloro-1-methyl-6-(4-methylphenyl)hydropyrazin-2-one

To a mixture of 2-(methylamino)-2-(4-methylphenyl)ethanenitrile, HClsalt (2 g, 10 mmol) in 1,2-dichlorobenzene (10 mL) was added (COCl)₂(6.5g, 50 mmol). The mixture was stirred at 90° C. for 1 h and cooled to 40°C. Four drops of DMF were added and the mixture was stirred at 125° C.for 3 h. The solvent was concentrated in vacuo and the residue waspurified by silica chromatography (1:5, EA:PE) to give 1.3 g (42%) ofthe title compound. ¹H NMR (400 MHz, CDCl₃): δ 2.47 (s, 3H), 3.34 (s,3H), 7.21 (d, J=10.8 Hz, 2H), 7.38 (d, J=10.8 Hz, 2H).

Preparation 16C:(tert-butoxy)-N-{1-[6-chloro-4-methyl-5-(4-methylphenyl)-3-oxo(4-hydropyrazin-2-yl)](4-piperidyl)}carboxamide

To a solution of3,5-dichloro-1-methyl-6-(4-methylphenyl)hydropyrazin-2-one (300 mg, 1.12mmol) in DMF (10 mL) was added (tert-butoxy)-N-(4-piperidyl)carboxamide(245 mg, 1.23 mmol) and DIEA (290 mg, 2.24 mmol). The mixture wasstirred at 120° C. for 3 h. DMF was concentrated in vacuo and theresidue partitioned between water and EA. The organic phase was washedwith brine, dried and concentrated to give 350 mg (73%) of the titleproduct. ¹H NMR (400 MHz, CDCl₃): δ 1.41-1.57 (m, 11H), 2.04-2.09 (m,2H), 2.43 (s, 3H), 3.07 (m, 2H), 3.19 (s, 3H), 3.70-3.76 (m, 1H), 4.53(s, 1H), 4.75 (d, J=13.6 Hz, 2H), 7.16 (d, J=10.8 Hz, 2H), 7.29 (d,J=10.8 Hz, 2H). [M+H] Calc'd for C₂₂H₂₉ClN₄O₃, 433; Found, 433.

Preparation 16D:{1-[6-(4-Cyano-phenyl)-4-methyl-3-oxo-5-p-tolyl-3,4-dihydro-pyrazin-2-yl]-piperidin-4-yl}-carbamicacid tert-butyl ester

To a solution of(tert-butoxy)-N-{1-[6-chloro-4-methyl-5-(4-methylphenyl)-3-oxo(4-hydropyrazin-2-yl)](4-piperidyl)}carboxamide(350 mg, 0.81 mmol) in DMF (10 mL) was added 4-cyanophenylboronic acid(238 mg, 1.62 mmol),[1,1′-Bis(di-tert-butylphosphino)ferrocene]dichloropalladium(II) (52 mg,0.08 mol) and K₂CO₃ (110 mg, 0.78 mmol) under N₂ atmosphere. The mixturewas stirred at 140° C. for 4 h. Water was added and the productextracted with EA (3×). The organics were combined, washed with brine,dried and concentrated. The residue was purified by columnchromatography (1:5, EA:PE) to give 200 mg (49%) of the title compound.[M+H] Calc'd for C₂₉H₃₃N₅O₃, 500; Found, 500.

Example 16:4-[6-(4-aminopiperidin-1-yl)-4-methyl-3-(4-methylphenyl)-5-oxo-4,5-dihydropyrazin-2-yl]benzonitrile

To a solution of{1-[6-(4-Cyano-phenyl)-4-methyl-3-oxo-5-p-tolyl-3,4-dihydro-pyrazin-2-yl]-piperidin-4-yl}-carbamicacid tert-butyl ester (200 mg, 0.4 mmol) in EA (10 mL) was added asolution of 4 N HCl in EA (10 mL). The mixture was stirred at RT for 2h. The solvent was concentrated in vacuo and the residue was purified bypreparative HPLC to give the product as the HCl salt (120 mg, 69%).¹H-NMR (CD₃OD, 400 MHz): δ 1.84-1.94 (m, 2H), 2.16-2.19 (m, 2H), 2.35(s, 3H), 3.24-3.30 (m, 5H), 3.49-3.54 (m, 1H), 4.88-4.93 (m, 2H), 7.16(d, J=6.0 Hz, 2H), 7.23 (d, J=5.6 Hz, 2H), 7.40 (d, J=6.4 Hz, 2H), 7.53(d, J=6.0 Hz, 2H). [M+H] Calc'd for C₂₄H₂₅N₅O, 400; Found, 400.

Example 17:4-[4-methyl-3-(4-methylphenyl)-5-oxo-6-{[(3S)-pyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin-2-yl]benzonitrile

The title compound was prepared as the HCl salt in 75% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.75-1.80 (m, 1H), 2.17-2.23 (m, 4H), 2.78-2.81(m, 1H), 3.02-3.07 (m, 1H), 3.17 (s, 3H), 3.35-3.45 (m, 3H), 3.63 (d,J=6.8 Hz, 2H), 7.09 (s, 4H), 7.41 (d, J=7.6 Hz, 2H), 7.51 (d, J=7.2 Hz,2H). [M+H] Calc'd for C₂₄H₂₅N₅O, 400; Found, 400.

Example 18:4-[4-methyl-3-(4-methylphenyl)-5-oxo-6-{[(3R)-pyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin-2-yl]benzonitrile

The title compound was prepared as the HCl salt in 70% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.75-1.80 (m, 1H), 2.12-2.23 (m, 4H), 2.78-2.83(m, 1H), 3.01-3.09 (m, 1H), 3.17 (s, 3H), 3.34-3.45 (m, 3H), 3.63 (d,J=6.8 Hz, 2H), 7.08 (s, 4H), 7.41 (d, J=7.6 Hz, 2H), 7.51 (d, J=7.2 Hz,2H). [M+H] Calc'd for C₂₄H₂₅N₅O, 400; Found, 400.

Example 19:5-[6-(4-aminopiperidin-1-yl)-4-methyl-3-(4-methylphenyl)-5-oxo-4,5-dihydropyrazin-2-yl]pyridine-2-carbonitrile

The title compound was prepared as the HCl salt in 60% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (CD₃OD, 400 MHz): δ 1.66-1.74 (m, 2H), 2.01-2.11 (m, 2H), 2.32 (s,3H), 2.90-3.03 (m, 2H), 3.23 (s, 3H), 3.40-3.51 (m, 1H), 4.80-4.85 (m,2H), 7.12 (d, J=7.6 Hz, 2H), 7.23 (d, J=8.0 Hz, 2H), 7.58 (d, J=6.8 Hz,1H), 7.72 (d, J=6.4 Hz, 1H), 8.41 (s, 1H). [M+H] Calc'd for C₂₃H₂₄N₆O,401; Found, 401.

Example 20:4-{4-methyl-6-[4-(methylamino)piperidin-1-yl]-3-(4-methylphenyl)-5-oxo-4,5-dihydropyrazin-2-yl}benzonitrile

The title compound was prepared as the HCl salt in 70% overall yieldaccording to the general procedure for the preparation of Example 16.¹H-NMR (CD₃OD, 400 MHz): δ 1.94-1.99 (m, 2H), 2.31-2.45 (m, 5H), 2.79(s, 3), 3.28-3.37 (m, 5H), 3.48-3.54 (m, 1H), 4.97-4.99 (m, 2H), 7.19(d, J=8.0 Hz, 2H), 7.26 (d, J=8.0 Hz, 2H), 7.46 (d, J=7.6 Hz, 2H), 7.58(d, J=8.0 Hz, 2H). [M+H] Calc'd for C₂₅H₂₇N₅O, 414; Found, 414.

Example 21:4-[6-(4-amino-4-methylpiperidin-1-yl)-4-methyl-3-(4-methylphenyl)-5-oxo-4,5-dihydropyrazin-2-yl]benzonitrile

The title compound was prepared as the HCl salt in 69% overall yieldaccording to the general procedure for the preparation of Example 16.¹H-NMR (DMSO-d₆, 400 MHz): δ 1.39 (s, 3H), 1.74-1.91 (m, 4H), 2.35 (s,3H), 3.13 (s, 3H), 3.46-3.51 (m, 2H), 4.28-4.32 (m, 2H), 7.19-7.29 (m,6H), 7.59 (d, J=8.0 Hz, 2H), 8.29 (br, 2H). [M+H] Calc'd for C₂₅H₂₇N₅O,414; Found, 414.

Example 22:4-[6-(3-aminopiperidin-1-yl)-4-methyl-3-(4-methylphenyl)-5-oxo-4,5-dihydropyrazin-2-yl]benzonitrile

The title compound was prepared in 43% overall yield according to thegeneral procedure for the preparation of Example 16. ¹H NMR (400 MHz,CD₃OD): δ 1.82-1.89 (m, 2H), 2.08-2.18 (m, 2H), 2.39 (s, 3H), 3.00-3.09(m, 2H), 3.29-3.33 (m, 4H), 3.57-3.62 (m, 1H), 4.34-4.92 (m, 1H), 7.15(d, J=8.1 Hz, 2H), 7.25 (d, J=8.1 Hz, 2H), 7.36 (d, J=8.4 Hz, 2H), 7.46(d, J=8.4 Hz, 2H). [M+H] Calc'd for C₂₄H₂₅N₅O, 400; Found, 400.

Example 23:4-[4-methyl-3-(4-methylphenyl)-6-{octahydro-1H-pyrrolo[3,4-c]pyridin-5-yl}-5-oxo-4,5-dihydropyrazin-2-yl]benzonitrile

The title compound was prepared in 64% overall yield according to thegeneral procedure for the preparation of Example 16. ¹H NMR (400 MHz,CD₃OD): δ 1.67-1.72 (m, 1H), 1.90-1.96 (m, 1H), 2.39 (s, 3H), 2.61-2.69(m, 2H), 3.22-3.61 (m, 9H), 4.35-4.44 (m, 2H), 7.15 (d, J=8.0 Hz, 2H),7.27 (d, J=7.6 Hz, 2H), 7.34 (d, J=8.4 Hz, 2H), 7.49 (d, J=8.4 Hz, 2H).[M+H] Calc'd for C₂₆H₂₇N₅O, 426; Found, 426.

Example 24:4-(6-{2,8-diazaspiro[4.5]decan-8-yl}-4-methyl-3-(4-methylphenyl)-5-oxo-4,5-dihydropyrazin-2-yl)benzonitrile

The title compound was prepared as the HCl salt in 45% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, DMSO-d₆): δ 1.66-1.69 (m, 4H), 1.84-1.89 (m, 2H), 2.34 (s,3H), 3.05-3.29 (m, 7H), 3.72-3.88 (m, 4H), 7.19-7.29 (m, 6H), 7.59 (d,J=8.4 Hz, 2H), 8.93-8.95 (br, 2H). [M+H] Calc'd for C₂₇H₂₉N₅O, 440;Found, 440.

Example 25:4-(6-{decahydropyrrolo[3,4-d]azepin-6-yl}-4-methyl-3-(4-methylphenyl)-5-oxo-4,5-dihydropyrazin-2-yl)benzonitrile

The title compound was prepared as the HCl salt in 64% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.89-2.07 (m, 4H), 2.39 (s, 3H), 2.74-2.76 (m,2H), 3.00-3.03 (m, 2H), 3.32 (s, 3H), 3.52-3.67 (m, 4H), 4.33-4.39 (m,2H), 7.15 (d, J=7.6 Hz, 2H), 7.27 (d, J=7.6 Hz, 2H), 7.34 (d, J=8.4 Hz,2H), 7.45 (d, J=8.4 Hz, 2H). [M+H] Calc'd for C₂₇H₂₉N₅O, 440; Found,440.

Example 26:4-[6-(4-aminopiperidin-1-yl)-4-ethyl-3-(4-methylphenyl)-5-oxo-4,5-dihydropyrazin-2-yl]benzonitrile

The title compound was prepared as the HCl salt in 89% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.15 (t, J=6.8 Hz, 3H), 1.85-1.95 (m, 2H),2.17-2.20 (m, 2H), 2.39 (s, 3H), 3.19-3.25 (t, J=8.0 Hz, 2H), 3.48-3.54(m, 1H), 3.86-3.90 (m, 2H), 4.87-4.90 (m, 2H), 7.22-7.28 (m, 4H),7.39-7.54 (m, 4H). [M+H] Calc'd for C₂₅H₂₇N₅O, 414; Found, 414.

Example 27:4-[6-(4-aminopiperidin-1-yl)-3-(4-ethylphenyl)-4-methyl-5-oxo-4,5-dihydropyrazin-2-yl]benzonitrile

The title compound was prepared as the HCl salt in 44% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.22 (t, J=7.2 Hz, 3H), 1.74-1.84 (m, 2H),2.09-2.13 (m, 2H), 2.69-2.75 (m, 2H), 3.03 (t, J=12.4 Hz, 2H), 3.30 (s,3H), 3.39-3.46 (m, 1H), 4.87-4.90 (m, 2H), 7.20 (d, J=8.0 Hz, 2H),7.31-7.36 (m, 4H), 7.47 (d, J=8.0 Hz, 2H). [M+H] Calc'd for C₂₅H₂₇N₅O,414; Found, 414.

Example 28:4-[3-(4-ethylphenyl)-4-methyl-5-oxo-6-{[(3S)-pyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin-2-yl]benzonitrile

The title compound was prepared as the HCl salt in 67% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.22 (t, J=7.6 Hz, 3H), 1.88-1.93 (m, 1H),2.28-2.34 (m, 1H), 263-2.68 (m, 2H), 2.90-2.94 (m, 1H), 3.15-3.19 (m,1H), 3.29 (s, 3H), 3.36-3.58 (m, 3H), 3.76 (d, J=7.2 Hz, 2H), 7.21-7.25(m, 4H), 7.54 (d, J=8.0 Hz, 2H), 7.64 (d, J=8.0 Hz, 2H). [M+H] Calc'dfor C₂₅H₂₇N₅O, 414; Found, 414.

Example 29:4-[3-(4-ethylphenyl)-4-methyl-5-oxo-6-{[(3R)-pyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin-2-yl]benzonitrile

The title compound was prepared as the HCl salt in 70% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.22 (t, J=7.6 Hz, 3H), 1.88-1.93 (m, 1H),2.28-2.34 (m, 1H), 263-2.68 (m, 2H), 2.90-2.94 (m, 1H), 3.15-3.19 (m,1H), 3.29 (s, 3H), 3.36-3.58 (m, 3H), 3.76 (d, J=7.2 Hz, 2H), 7.21-7.25(m, 4H), 7.54 (d, J=8.0 Hz, 2H), 7.64 (d, J=8.0 Hz, 2H). [M+H] Calc'dfor C₂₅H₂₇N₅O, 414; Found, 414.

Example 30:4-[6-(4-aminopiperidin-1-yl)-3-(4-methoxyphenyl)-4-methyl-5-oxo-4,5-dihydropyrazin-2-yl]benzonitrile

The title compound was prepared as the HCl salt in 79% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.74-1.84 (m, 2H), 2.09-2.13 (m, 2H), 3.02 (t,J=12.0 Hz, 2H), 3.30 (s, 3H), 3.39-3.46 (m, 1H), 3.85 (s, 3H), 4.87-4.90(m, 2H), 7.00 (d, J=4.8 Hz, 2H), 7.20 (d, J=8.0 Hz, 2H), 7.36 (d, J=8.4Hz, 2H), 7.49 (d, J=8.4 Hz, 2H). [M+H] Calc'd for C₂₄H₂₅N₅O₂, 416;Found, 416.

Example 31:4-[3-(4-methoxyphenyl)-4-methyl-5-oxo-6-{[(3S)-pyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin-2-yl]benzonitrile

The title compound was prepared as the HCl salt in 80% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.88-1.93 (m, 1H), 2.30-2.34 (m, 1H), 2.90-2.94(m, 1H), 3.15-3.19 (m, 1H), 3.31 (s, 3H), 3.36-3.40 (m, 1H), 3.47-3.58(m, 2H), 3.76 (d, J=7.2 Hz, 2H), 3.79 (s, 3H), 6.93 (d, J=8.0 Hz, 2H),7.24 (d, J=8.0 Hz, 2H), 7.54 (d, J=7.6 Hz, 2H), 7.66 (d, J=7.6 Hz, 2H).[M+H] Calc'd for C₂₄H₂₅N₅O₂,416; Found, 416.

Example 32:4-[3-(4-methoxyphenyl)-4-methyl-5-oxo-6-{[(3R)-pyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin-2-yl]benzonitrile

The title compound was prepared as the HCl salt in 57% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.88-1.93 (m, 1H), 2.30-2.34 (m, 1H), 2.90-2.94(m, 1H), 3.15-3.19 (m, 1H), 3.31 (s, 3H), 3.36-3.40 (m, 1H), 3.47-3.58(m, 2H), 3.76 (d, J=7.2 Hz, 2H), 3.79 (s, 3H), 6.93 (d, J=8.0 Hz, 2H),7.24 (d, J=8.0 Hz, 2H), 7.54 (d, J=7.6 Hz, 2H), 7.66 (d, J=7.6 Hz, 2H).[M+H] Calc'd for C₂₄H₂₅N₅O₂, 416; Found, 416.

Example 33:4-[6-(4-aminopiperidin-1-yl)-3-(4-cyclopropylphenyl)-4-methyl-5-oxo-4,5-dihydropyrazin-2-yl]benzonitrile

The title compound was prepared as the HCl salt in 87% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 0.73-0.77 (m, 2H), 1.03-1.06 (m, 2H), 1.74-1.84(m, 2H), 1.94-1.99 (m, 1H), 2.09-2.13 (m, 2H), 3.02 (t, J=12.0 Hz, 2H),3.30 (s, 3H), 3.39-3.46 (m, 1H), 4.87-4.90 (m, 2H), 7.16 (s, 4H), 7.35(d, J=8.0 Hz, 2H), 7.48 (d, J=8.0 Hz, 2H). [M+H] Calc'd for C₂₆H₂₇N₅O,426; Found, 426.

Example 34:4-[3-(4-cyclopropylphenyl)-4-methyl-5-oxo-6-{[(3S)-pyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin-2-yl]benzonitrile

The title compound was prepared as the HCl salt in 55% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 0.67-0.73 (m, 2H), 0.99-1.04 (m, 2H), 1.88-1.93(m, 2H), 2.30-2.34 (m, 1H), 2.90-2.94 (m, 1H), 3.15-3.19 (m, 1H), 3.29(s, 3H), 3.36-3.40 (m, 1H), 3.47-3.58 (m, 2H), 3.76 (d, J=7.2 Hz, 2H),7.08 (d, J=8.0 Hz, 2H), 7.19 (d, J=8.0 Hz, 2H), 7.53 (d, J=7.6 Hz, 2H),7.65 (d, J=7.6 Hz, 2H). [M+H] Calc'd for C₂₆H₂₇N₅O 426; Found, 426.

Example 35:4-[3-(4-cyclopropylphenyl)-4-methyl-5-oxo-6-{[(3R)-pyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin-2-yl]benzonitrile

The title compound was prepared as the HCl salt in 45% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 0.67-0.73 (m, 2H), 0.99-1.04 (m, 2H), 1.88-1.93(m, 2H), 2.30-2.34 (m, 1H), 2.90-2.94 (m, 1H), 3.15-3.19 (m, 1H), 3.29(s, 3H), 3.36-3.40 (m, 1H), 3.47-3.58 (m, 2H), 3.76 (d, J=7.2 Hz, 2H),7.08 (d, J=8.0 Hz, 2H), 7.19 (d, J=8.0 Hz, 2H), 7.53 (d, J=7.6 Hz, 2H),7.65 (d, J=7.6 Hz, 2H). [M+H] Calc'd for C₂₆H₂₇N₅O 426; Found, 426.

Example 36:4-[6-(4-amino-piperidin-1-yl)-4-(2-methoxy-ethyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]-benzonitrile

The title compound was prepared as the HCl salt in 72% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, DMSO-d₆): δ 1.62-1.65 (m, 2H), 1.99-2.01 (m, 2H), 2.34 (s,3H), 2.96 (t, J=12.4 Hz, 2H), 3.05 (s, 3H), 3.25-3.35 (m, 1H), 3.37 (t,J=6.4 Hz, 2H), 3.87 (t, J=6.4 Hz, 2H), 4.72 (d, J=13.6 Hz, 2H),7.21-7.27 (m, 6H), 7.58 (d, J=8.4 Hz, 2H), 8.17 (br, 3H). [M+H] Calc'dfor C₂₆H₂₉N₅O₂, 444; Found, 444.

Example 37:4-[6-(4-amino-piperidin-1-yl)-4-(2-hydroxy-ethyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]-benzonitrile

To a solution of{1-[6-(4-cyano-phenyl)-4-(2-methoxy-ethyl)-3-oxo-5-p-tolyl-3,4-dihydro-pyrazin-2-yl]-piperidin-4-yl}-carbamicacid tert-butyl ester (500 mg, 0.92 mmol) in DCM was added a 1N solutionof BBr₃ in DCM (1.84 mL, 1.84 mmol) at −78° C., then stirred for 1 h atRT. Aq NaHCO₃ was added until the pH was adjusted to pH=7-8. The organiclayer was dried, concentrated and purified by HPLC to give the titlecompound as the HCl salt (101 mg, 26%). ¹H NMR (400 MHz, DMSO-d₆): δ1.65-1.68 (m, 2H), 2.00-2.03 (m, 2H), 2.35 (s, 3H), 2.96 (t, J=12.0 Hz,2H), 3.30-3.40 (m, 1H), 3.43 (t, J=6.4 Hz, 2H), 3.78 (t, J=6.4 Hz, 2H),4.73 (d, J=13.2 Hz, 2H), 7.24-7.27 (m, 6H), 7.58 (d, J=8.4 Hz, 2H), 8.29(br, 3H). [M+H] Calc'd for C₂₅H₂₇N₅O₂, 430; Found, 430.

Example 38:4-[6-(4-amino-piperidin-1-yl)-3-(4-cyclopropyl-phenyl)-4-(2-hydroxy-ethyl)-5-oxo-4,5-dihydro-pyrazin-2-yl]-benzonitrile

The title compound was prepared as the HCl salt in 57% overall yieldaccording to the general procedure for the preparation of Example 37. ¹HNMR (400 MHz, DMSO-d₆): δ 0.71-0.72 (m, 2H), 0.99-1.01 (m, 2H),1.60-1.64 (m, 2H), 1.95-2.00 (m, 3H), 2.94 (t, J=12.0 Hz, 2H), 3.24-3.34(m, 1H), 3.42 (t, J=6.8 Hz, 2H), 3.77 (t, J=6.4 Hz, 2H), 4.72 (d, J=13.2Hz, 2H), 7.10-7.25 (m, 6H), 7.59 (d, J=8.4 Hz, 2H), 8.09 (br, 3H). [M+H]Calc'd for C₂₇H₂₉N₅O₂, 456; Found, 456.

Example 39:4-[6-(4-amino-piperidin-1-yl)-4-(3-methoxy-propyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]-benzonitrile

The title compound was prepared as the HCl salt in 61% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, DMSO-d₆): δ 1.65-1.68 (m, 4H), 1.98-2.01 (m, 2H), 2.34 (s,3H), 2.95 (t, J=12.0 Hz, 2H), 3.03 (s, 3H), 3.13 (t, J=6.0 Hz, 2H),3.24-3.34 (m, 1H), 3.70-3.74 (m, 2H), 4.73 (d, J=12.4 Hz, 2H), 7.24-7.28(m, 6H), 7.59 (d, J=8.4 Hz, 2H), 8.11 (br, 3H). [M+H] Calc'd forC₂₇H₃₁N₅O₂, 458; Found, 458.

Example 40:4-[6-(4-amino-piperidin-1-yl)-4-(3-hydroxy-propyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]-benzonitrile

The title compound was prepared as the HCl salt in 23% overall yieldaccording to the general procedure for the preparation of Example 37. ¹HNMR (400 MHz, DMSO-d₆): δ 1.58-1.65 (m, 4H), 1.99-2.02 (m, 2H), 2.34 (s,3H), 2.95 (t, J=12.0 Hz, 2H), 3.19 (t, J=6.4 Hz, 2H), 3.24-3.34 (m, 1H),3.69-3.73 (m, 2H), 4.73 (d, J=13.2 Hz, 2H), 7.22-7.29 (m, 6H), 7.58 (d,J=8.4 Hz, 2H), 8.22 (br, 3H). [M+H] Calc'd for C₂₆H₂₉N₅O₂, 444; Found,444.

Example 41:4-[6-(4-amino-piperidin-1-yl)-4-(2-methoxy-ethyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]-2-fluoro-benzonitrile

The title compound was prepared as the HCl salt in 80% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, DMSO-d₆): δ 1.60-1.63 (m, 2H), 1.99-2.01 (m, 2H), 2.36 (s,3H), 2.96 (t, J=12.0 Hz, 2H), 3.05 (s, 3H), 3.25-3.35 (m, 1H), 3.37 (t,J=6.4 Hz, 2H), 3.88 (t, J=6.4 Hz, 2H), 4.73 (d, J=12.8 Hz, 2H), 7.03 (d,J=8.4 Hz, 1H), 7.13 (d, J=11.6 Hz, 1H), 7.24-7.30 (m, 4H), 7.66 (t,J=7.8 Hz, 1H), 8.06 (br, 3H). [M+H] Calc'd for C₂₆H₂₈FN₅O₂, 462; Found,462.

Example 42:4-[6-(4-amino-piperidin-1-yl)-4-(2-hydroxy-ethyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]-2-fluoro-benzonitrile

The title compound was prepared as the HCl salt in 50% overall yieldaccording to the general procedure for the preparation of Example 37. ¹HNMR (400 MHz, DMSO-d₆): δ 1.62-1.66 (m, 2H), 1.99-2.02 (m, 2H), 2.36 (s,3H), 2.96 (t, J=11.6 Hz, 2H), 3.30-3.40 (m, 1H), 3.42 (t, J=6.6 Hz, 2H),3.77 (t, J=6.6 Hz, 2H), 4.73 (d, J=12.8 Hz, 2H), 7.04 (d, J=8.0 Hz, 1H),7.11 (d, J=12.0 Hz, 1H), 7.24-7.29 (m, 4H), 7.66 (t, J=7.8 Hz, 1H), 8.15(br, 3H). [M+H] Calc'd for C₂₅H₂₆FN₅O₂, 448; Found, 448.

Example 43:4-(6-{[((3S)-pyrrolidin-3-yl)methyl]amino}-4-(2-methoxyethyl)-3-(4-methylphenyl)-5-oxo(4-hydropyrazin-2-yl))-2-fluorobenzenecarbonitrile

The title compound was prepared as the HCl salt in 44% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, DMSO-d₆): δ 1.68-1.71 (m, 1H), 1.99-2.01 (m, 1H), 2.35 (s,3H), 2.68-2.72 (m, 1H), 2.91-2.92 (m, 1H), 3.05 (s, 3H), 3.11-3.25 (m,3H), 3.34-3.47 (m, 4H), 3.88 (t, J=6.4 Hz, 2H), 7.08-7.15 (m, 2H),7.24-7.26 (m, 4H), 7.64 (t, J=7.6 Hz, 1H), 7.80 (s, 1H), 9.24 (br, 2H).[M+H] Calc'd for C₂₆H₂₈FN₅O₂, 462; Found, 462.

Example 44:4-(6-{[((3S)-pyrrolidin-3-yl)methyl]amino}-4-(2-hydroxyethyl)-3-(4-methylphenyl)-5-oxo(4-hydropyrazin-2-yl))-2-fluorobenzenecarbonitrile

The title compound was prepared as the HCl salt in 32% overall yieldaccording to the general procedure for the preparation of Example 37. 1HNMR (400 MHz, DMSO-d₆): δ 1.69-1.71 (m, 1H), 1.99-2.00 (m, 1H), 2.35 (s,3H), 2.66-2.71 (m, 1H), 2.91-2.92 (m, 1H), 3.11-3.25 (m, 3H), 3.40-3.46(m, 4H), 3.77 (t, J=6.6 Hz, 2H), 7.06-7.10 (m, 2H), 7.13-7.25 (m, 4H),7.64 (t, J=7.6 Hz, 1H), 7.79 (s, 1H), 9.16 (br, 2H). [M+H] Calc'd forC₂₅H₂₆FN₅O₂, 448; Found, 448.

Example 45:4-[6-(4-aminopiperidyl)-4-(3-methoxypropyl)-3-(4-methylphenyl)-5-oxo(4-hydropyrazin-2-yl)]-2-fluorobenzenecarbonitrile

The title compound was prepared as the HCl salt in 75% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, DMSO-d₆): δ 1.63-1.69 (m, 4H), 2.00-2.03 (m, 2H), 2.36 (s,3H), 2.96 (t, J=12.0 Hz, 2H), 3.03 (s, 3H), 3.13 (t, J=6.0 Hz, 2H),3.20-3.30 (m, 1H), 3.70-3.74 (m, 2H), 4.73 (d, J=13.6 Hz, 2H), 7.06 (d,J=8.0 Hz, 1H), 7.14 (d, J=10.4 Hz, 1H), 7.25-7.30 (m, 4H), 7.66 (d,J=8.0 Hz, 1H), 8.24 (br, 3H). [M+H] Calc'd for C₂₇H₃₀FN₅O₂, 476; Found,476.

Example 46:4-[6-(4-aminopiperidyl)-4-(3-hydroxypropyl)-3-(4-methylphenyl)-5-oxo(4-hydropyrazin-2-yl)]-2-fluorobenzenecarbonitrile

The title compound was prepared as the HCl salt in 28% overall yieldaccording to the general procedure for the preparation of Example 37. ¹HNMR (300 MHz, DMSO-d₆): δ 1.57-1.67 (m, 4H), 1.99-2.03 (m, 2H), 2.36 (s,3H), 2.95 (t, J=12.0 Hz, 2H), 3.19 (t, J=6.3 Hz, 2H), 3.25-3.35 (m, 1H),3.69-3.74 (m, 2H), 4.73 (d, J=12.6 Hz, 2H), 7.04-7.30 (m, 6H), 7.66 (d,J=7.6 Hz, 1H), 8.24 (br, 3H). [M+H] Calc'd for C₂₆H₂₈FN₅O₂, 462; Found,462.

Example 47:4-[6-(4-amino-piperidin-1-yl)-4-methyl-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]-2-fluoro-benzonitrile

The title compound was prepared as the HCl salt in 51% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.62-1.71 (m, 2H), 1.97-2.01 (m, 2H), 2.31 (s,3H), 2.91 (t, J=12.0 Hz, 2H), 3.12 (s, 3H), 3.28-3.33 (m, 1H), 4.87-4.90(m, 2H), 6.99-7.09 (m, 4H), 7.20-7.36 (m, 3H). [M+H] Calc'd forC₂₄H₂₄FN₅O, 418; Found, 418.

Example 48:4-[6-(4-amino-piperidin-1-yl)-4-methyl-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]-3-fluoro-benzonitrile

The title compound was prepared as the HCl salt in 43% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.68-1.77 (m, 2H), 2.01-2.03 (m, 2H), 2.22 (s,3H), 3.05 (t, J=12.0 Hz, 2H), 3.17 (s, 3H), 3.34-3.39 (m, 1H), 4.72-4.75(m, 2H), 7.02-7.09 (m, 4H), 7.29-7.36 (m, 3H). [M+H] Calc'd forC₂₄H₂₄FN₅O, 418; Found, 418.

Example 49:4-[6-(4-amino-piperidin-1-yl)-4-methyl-3-(6-methyl-pyridin-3-yl)-5-oxo-4,5-dihydro-pyrazin-2-yl]-benzonitrile

The title compound was prepared as the HCl salt in 40% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.90-1.97 (m, 2H), 2.19-2.23 (m, 2H), 2.85 (s,3H), 3.27-3.30 (m, 2H), 3.37 (s, 3H), 3.55-3.57 (m, 1H), 4.72-4.75 (m,2H), 7.51 (d, J=8.0 Hz, 2H), 7.66 (d, J=8.0 Hz, 2H), 8.029 (d, J=8.4 Hz,1H), 8.60 (d, J=8.4 Hz, 1H), 8.84 (s, 1H). [M+H] Calc'd for C₂₃H₂₄N₆O,401; Found, 401.

Example 50:4-[6-(4-amino-piperidin-1-yl)-3-(4-cyclopropyl-phenyl)-4-methyl-5-oxo-4,5-dihydro-pyrazin-2-yl]-2-fluoro-benzonitrile

The title compound was prepared as the HCl salt in 58% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 0.62-0.66 (m, 2H), 0.93-0.97 (m, 2H), 1.66-1.75(m, 2H), 1.83-1.88 (m, 1H), 2.01-2.04 (m, 2H), 2.98 (t, J=12.0 Hz, 2H),3.16 (s, 3H), 3.32-3.37 (m, 1H), 4.72-4.75 (m, 2H), 7.01-7.07 (m, 6H),7.36-7.39 (m, 1H). [M+H] Calc'd for C₂₆H₂₆FN₅O, 444; Found, 444.

Example 51:5-[6-(4-amino-piperidin-1-yl)-3-(4-methoxy-phenyl)-4-methyl-5-oxo-4,5-dihydro-pyrazin-2-yl]-pyridine-2-carbonitrile

The title compound was prepared as the HCl salt in 25% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.81-1.91 (m, 2H), 2.06-2.19 (m, 2H), 3.16-3.22(m, 2H), 3.22 (s, 3H), 3.47-3.53 (m, 1H), 3.86 (s, 3H), 4.72-4.75 (m,2H), 7.04 (d, J=8.4 Hz, 2H), 7.27 (d, J=8.4 Hz, 2H), 7.73 (d, J=8.4 Hz,1H), 7.84 (d, J=8.4 Hz, 1H), 8.50 (s, 1H). [M+H] Calc'd for C₂₃H₂₄N₆O₂,417; Found, 417.

Example 52:4-[6-(4-amino-piperidin-1-yl)-3-(4-methoxy-phenyl)-4-methyl-5-oxo-4,5-dihydro-pyrazin-2-yl]-2-fluoro-benzonitrile

The title compound was prepared as the HCl salt in 65% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.73-1.83 (m, 2H), 2.11-2.14 (m, 2H), 3.00-3.06(m, 2H), 3.34 (s, 3H), 3.38-3.46 (m, 1H), 3.86 (s, 3H), 4.80-4.90 (m,2H), 7.04-7.24 (m, 6H), 7.46-7.50 (m, 1H). [M+H] Calc'd for C₂₄H₂₄FN₅O₂,434; Found, 434.

Example 53:5-[6-(4-amino-piperidin-1-yl)-3-(4-cyclopropyl-phenyl)-4-methyl-5-oxo-4,5-dihydro-pyrazin-2-yl]-pyridine-2-carbonitrile

The title compound was prepared as the HCl salt in 65% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 0.64-0.67 (m, 2H), 0.92-0.98 (m, 2H), 1.64-1.73(m, 2H), 1.83-1.92 (m, 1H), 2.01-2.04 (m, 2H), 2.92-2.98 (m, 2H), 3.18(s, 3H), 3.32-3.39 (m, 1H), 4.72-4.75 (m, 2H), 7.08-7.16 (m, 4H),7.53-7.65 (m, 2H), 8.33 (s, 1H). [M+H] Calc'd for C₂₅H₂₆N₆O, 427; Found,427.

Example 54:5-{3-(4-methoxy-phenyl)-4-methyl-5-oxo-6-[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro-pyrazin-2-yl}-pyridine-2-carbonitrile

The title compound was prepared as the HCl salt in 55% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.88-1.93 (m, 1H), 2.24-2.28 (m, 1H), 2.87-2.94(m, 1H), 3.11-3.16 (m, 1H), 3.28-3.35 (m, 4H), 3.42-3.52 (m, 2H), 3.76(d, J=7.2 Hz, 2H), 3.86 (s, 3H), 7.01 (d, J=8.4 Hz, 2H), 7.50 (d, J=8.4Hz, 2H), 7.72 (d, J=8.0 Hz, 1H), 8.30 (d, J=8.0 Hz, 1H), 8.54 (s, 1H).[M+H] Calc'd for C₂₃H₂₄N₆O2, 417; Found, 417.

Example 55:2-fluoro-4-{3-(4-methoxy-phenyl)-4-methyl-5-oxo-6-[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro-pyrazin-2-yl}-benzonitrile

The title compound was prepared as the HCl salt in 52% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.88-1.97 (m, 1H), 2.29-2.37 (m, 1H), 2.91-2.97(m, 1H), 3.17-3.22 (m, 1H), 3.33-3.41 (m, 4H), 3.47-3.59 (m, 2H),3.78-3.82 (m, 5H), 6.97 (d, J=8.4 Hz, 2H), 7.27-7.49 (m, 4H), 7.65-7.69(m, 1H). [M+H] Calc'd for C₂₄H₂₄FN₅O₂, 434; Found, 434.

Example 56:5-{3-(4-ethyl-phenyl)-4-methyl-5-oxo-6-[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro-pyrazin-2-yl}-pyridine-2-carbonitrile

The title compound was prepared as the HCl salt in 70% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.15 (t, J=7.6 Hz, 3H), 1.74-1.82 (m, 1H),2.01-2.15 (m, 1H), 2.62 (q, J=7.6 Hz, 2H), 2.73-2.79 (m, 1H), 2.97-3.02(m, 1H), 3.14-3.34 (m, 4H), 3.27-3.37 (m, 2H), 3.48-3.58 (m, 2H), 7.12(d, J=8.4 Hz, 2H), 7.21 (d, J=8.4 Hz, 2H), 7.53 (d, J=8.0 Hz, 1H),7.63-7.66 (m, 1H), 8.37 (s, 1H). [M+H] Calc'd for C₂₄H₂₆N₆O, 415; Found,415.

Example 57:4-{3-(4-ethyl-phenyl)-4-methyl-5-oxo-6-[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro-pyrazin-2-yl}-2-fluoro-benzonitrile

The title compound was prepared as the HCl salt in 66% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.24 (t, J=7.6 Hz, 3H), 1.86-1.95 (m, 1H),2.27-2.35 (m, 1H), 2.69 (q, J=7.6 Hz, 2H), 2.89-2.96 (m, 1H), 3.15-3.21(m, 1H), 3.14-3.34 (m, 4H), 3.39-3.55 (m, 2H), 3.57-3.77 (m, 2H),7.24-7.39 (m, 6H), 7.62-7.66 (m, 1H). [M+H] Calc'd for C₂₅H₂₆FN₅O, 432;Found, 432.

Example 58:5-{3-(4-cyclopropyl-phenyl)-4-methyl-5-oxo-6-[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro-pyrazin-2-yl}-pyridine-2-carbonitrile

The title compound was prepared as the HCl salt in 47% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 0.70-0.74 (m, 2H), 1.02-1.07 (m, 2H), 1.88-1.97(m, 2H), 2.29-2.35 (m, 1H), 2.91-2.97 (m, 1H), 3.16-3.19 (m, 1H),3.30-3.38 (m, 4H), 3.47-3.59 (m, 2H), 3.75-3.78 (m, 2H), 7.13 (d, J=8.0Hz, 2H), 7.20-7.24 (m, 2H), 7.79 (d, J=8.0 Hz, 1H), 7.96-7.99 (m, 1H),8.59 (s, 1H). [M+H] Calc'd for C₂₅H₂₆N₆O, 427; Found, 427.

Example 59:4-{3-(4-cyclopropyl-phenyl)-4-methyl-5-oxo-6-[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro-pyrazin-2-yl}-2-fluoro-benzonitrile

The title compound was prepared as the HCl salt in 47% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 0.70-0.74 (m, 2H), 1.02-1.07 (m, 2H), 1.85-1.97(m, 2H), 2.27-2.35 (m, 1H), 2.89-2.95 (m, 1H), 3.16-3.19 (m, 1H),3.30-3.38 (m, 4H), 3.47-3.59 (m, 2H), 3.75-3.78 (m, 2H), 7.13 (d, J=8.0Hz, 2H), 7.18-7.22 (m, 2H), 7.29 (d, J=8.4 Hz, 1H), 7.37 (d, J=8.4 Hz,1H), 7.61-7.65 (m, 1H). [M+H] Calc'd for C₂₆H₂₆FN₅O, 444; Found, 444.

Example 60:4-[6-(4-amino-piperidin-1-yl)-4-methyl-3-(1-methyl-1H-benzoimidazol-5-yl)-5-oxo-4,5-dihydro-pyrazin-2-yl]-2-fluoro-benzonitrile

The title compound was prepared as the HCl salt in 43% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.68-1.77 (m, 2H), 2.03-2.06 (m, 2H), 3.02 (t,J=12.0 Hz, 2H), 3.16 (s, 3H), 3.34-3.40 (m, 1H), 4.09 (s, 3H), 4.80-4.90(m, 2H), 6.97 (d, J=8.4 Hz, 1H), 7.16 (d, J=10.8 Hz, 1H), 7.32 (d, J=8.4Hz, 1H), 7.60 (d, J=8.4 Hz, 1H), 7.79 (s, 1H), 7.96 (d, J=8.4 Hz, 1H),9.42 (s, 1H). [M+H] Calc'd for C₂₅H₂₄FN₇O, 458; Found, 458.

Example 61:4-{3-(4-cyclopropylmethoxy-phenyl)-4-methyl-5-oxo-6-[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro-pyrazin-2-yl}-2-fluoro-benzonitrile

The title compound was prepared as the HCl salt in 60% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 0.34-0.38 (m, 2H), 0.61-0.66 (m, 2H), 1.24-1.27(m, 1H), 1.85-1.97 (m, 1H), 2.27-2.35 (m, 1H), 2.89-2.95 (m, 1H),3.14-3.19 (m, 1H), 3.30-3.38 (m, 4H), 3.47-3.59 (m, 2H), 3.75 (d, J=7.6Hz, 2H), 3.85 (d, J=7.6 Hz, 2H), 6.95 (d, J=8.4 Hz, 2H), 7.22-7.24 (m,2H), 7.29 (d, J=8.4 Hz, 1H), 7.39 (d, J=8.4 Hz, 1H), 7.65 (t, J=7.6 Hz,1H). [M+H] Calc'd for C₂₇H₂₈FN₅O₂, 474; Found, 474.

Example 62:4-{3-(4-ethoxy-phenyl)-4-methyl-5-oxo-6-[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro-pyrazin-2-yl}-2-fluoro-benzonitrile

The title compound was prepared as the HCl salt in 60% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.28 (t, J=7.2 Hz, 3H), 1.75-1.80 (m, 1H),2.15-2.20 (m, 1H), 2.77-2.82 (m, 1H), 3.01-3.06 (m, 1H), 3.18-3.26 (m,4H), 3.32-3.43 (m, 2H), 3.62 (d, J=7.6 Hz, 2H), 3.94 (d, J=7.6 Hz, 2H),6.83 (d, J=8.4 Hz, 2H), 7.09-7.11 (m, 2H), 7.17 (d, J=8.4 Hz, 1H), 7.25(d, J=8.4 Hz, 1H), 7.51 (t, J=7.6 Hz, 1H). [M+H] Calc'd for C₂₅H₂₆FN₅O₂,448; Found, 448.

Example 63:2-fluoro-4-[4-methyl-5-oxo-6-R[((3S)-pyrrolidin-3-ylmethyl)-amino]-3-(4-trifluoromethoxy-phenyl)-4,5-dihydro-pyrazin-2-yl]-benzonitrile

The title compound was prepared as the HCl salt in 60% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.85-1.97 (m, 1H), 2.27-2.35 (m, 1H), 2.89-2.95(m, 1H), 3.14-3.19 (m, 1H), 3.30-3.38 (m, 4H), 3.47-3.59 (m, 2H), 3.75(d, J=7.6 Hz, 2H), 7.24-7.53 (m, 6H), 7.63 (t, J=7.6 Hz, 1H). [M+H]Calc'd for C₂₄H₂₁F₄N₅O₂, 488; Found, 488.

Example 64:2-Fluoro-4-[4-(3-hydroxy-propyl)-6-(4-methylamino-piperidin-1-yl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]-benzonitrile

The title compound was prepared as the HCl salt in 20% overall yieldaccording to the general procedure for the preparation of Example 37. ¹HNMR (300 MHz, CD₃OD): δ 1.76-1.86 (m, 4H), 2.23-2.26 (m, 2H), 2.38 (s,3H), 2.75 (s, 3H), 3.14-3.15 (m, 2H), 3.38-3.40 (m, 3H), 3.87-3.90 (m,2H), 4.91-4.96 (m, 2H), 7.15-7.29 (m, 6H), 7.48-7.49 (m, 1H). [M+H]Calc'd for C₂₇H₃₀FN₅O₂, 476; Found, 476.

Example 65:2-Fluoro-4-[4-(2-hydroxy-ethyl)-6-(4-methylamino-piperidin-1-yl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]-benzonitrile

The title compound was prepared as the HCl salt in 37.5% overall yieldaccording to the general procedure for the preparation of Example 37. ¹HNMR (300 MHz, CD₃OD): δ 1.70-1.75 (m, 2H), 2.15-2.21 (m, 2H), 2.42 (s,3H), 2.74 (s, 3H), 2.99 (t, J=12.3 Hz, 2H), 3.29-3.30 (m, 1H), 3.64 (t,J=6.3 Hz, 2H), 4.00 (t, J=6.3 Hz, 2H), 4.87-4.88 (m, 2H), 7.12 (t,J=10.5 Hz, 2H), 7.22-7.32 (m, 4H), 7.45 (t, J=7.5 Hz, 1H). [M+H] Calc'dfor C₂₆H₂₈FN₅O₂, 462; Found, 462.

Example 66:4-[6-(4-Amino-piperidin-1-yl)-4-methyl-3-(6-methyl-pyridin-3-yl)-5-oxo-4,5-dihydro-pyrazin-2-yl]-2-fluoro-benzonitrile

The title compound was prepared as the HCl salt in 45% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.82-1.92 (m, 2H), 2.16-2.19 (m, 2H), 2.87 (s,3H), 3.18-3.24 (m, 2H), 3.32 (s, 3H), 3.49-3.55 (m, 1H), 4.86-4.88 (m,2H), 7.15 (d, J=8.0 Hz, 1H), 7.45 (d, J=8.0 Hz, 1H), 7.62 (d, J=8.4 Hz,1H), 8.04 (d, J=8.4 Hz, 1H), 8.61 (d, J=8.4 Hz, 1H), 8.85 (s, 1H). [M+H]Calc'd for C₂₃H₂₃FN₆O, 419; Found, 419

Preparation 67A: 6-chloro-1-methylpyridin-2(1H)-one

To a mixture of 6-chloropyridin-2-ol (10.0 g, 77.5 mmol) and K₂CO₃ (37.4g, 271.3 mmol) in acetone (100 mL) was added iodomethane (38.5 g, 271.3mmol) and the reaction was heated to reflux overnight. The reactionmixture was cooled to RT and the insoluble materials filtered. Thesolvent was concentrated in vacuo and the residue was partitionedbetween H₂O and DCM. The organic layer was dried and concentrated. Thesolids were slurried in diethyl ether at RT for 30 min, filtered, washedwith ether and dried to give 6.5 g (59%) of the title compound. ¹H NMR(400 MHz, MeOD-d₄): δ 3.73 (s, 3H), 6.51-6.59 (m, 2H), 7.45-7.49 (m,1H). [M+H] Calc'd for C₆H₆ClNO, 144; Found, 144.

Preparation 67B: 6-(4-methoxyphenyl)-1-methylpyridin-2(1H)-one

A mixture of 6-chloro-1-methylpyridin-2(1H)-one (520 mg, 3.64 mmol) intoluene/EtOH/H₂O (5 mL/5 mL/7.5 mL), 4-methoxyphenylboronic acid (553mg, 3.64 mmol), Pd(PPh₃)₄ (42 mg, 0.036 mmol), Na₂CO₃ (1157 mg, 10.92mmol) and LiCl.H₂O (218 mg, 3.64 mmol) was flushed with nitrogen andstirred overnight at 120° C. Water was added and the reaction wasextracted with EA (3×). The organics were combined, washed with brine,dried and concentrated. The residue was purified by silicachromatography (1:5, EA:PE) to give 0.55 g (70%) of the title compound.¹H NMR (400 MHz, CDCl₃): δ 3.39 (s, 3H), 3.86 (s, 3H), 6.07 (dd, J=1.2,6.8 Hz, 1H), 6.56 (dd, J=1.2, 9.2 Hz, 1H), 6.97-6.99 (m, 2H), 7.27-7.28(m, 2H), 7.31-7.35 (m, 1H). [M+H] Calc'd for C₁₃H₁₃NO₂, 216; Found, 216.

Preparation 67C:3,5-dibromo-6-(4-methoxyphenyl)-1-methylpyridin-2(1H)-one

To a solution of 6-(4-methoxyphenyl)-1-methylpyridin-2(1H)-one (0.55 g,2.42 mmol) in AcOH (10 mL) was added bromine (0.78 g, 4.84 mmol)dropwise, and the mixture was stirred at RT overnight. Water was addedand the reaction was extracted with DCM (3×). The organics werecombined, washed with brine, dried and concentrated to give 0.60 g (67%)of the title compound. ¹H NMR (400 MHz, CDCl₃): δ 3.34 (s, 3H), 3.88 (s,3H), 7.02 (d, J=8.8 Hz, 2H), 7.15 (d, J=8.4 Hz, 2H), 7.95 (s, 1H).

Preparation 67D:tert-butyl-1-(5-bromo-6-(4-methoxyphenyl)-1-methyl-2-oxo-1,2-dihydropyridin-3-yl)piperidin-4-ylcarbamate

A mixture of 3,5-dibromo-6-(4-methoxyphenyl)-1-methylpyridin-2(1H)-one(0.60 g, 1.62 mmol), (0.33 g, 1.62 mmol), Pd(OAc)₂ (36 mg, 0.16 mol),Cs₂CO₃ (5.3 g, 16.2 mmol) and BINAP (100 mg, 0.16 mmol) was flushed withnitrogen and stirred at 120° C. overnight. The reaction mixture wascooled to RT and the insoluble materials filtered. The solvent wasconcentrated in vacuo and the residue was purified by silicachromatography (1:1, EA:PE) to give 0.65 g (82%) of the title compound.¹H NMR (400 MHz, CDCl₃): δ 1.43 (s, 9H), 1.57-1.59 (m, 2H), 2.05-2.08(m, 2H), 2.63-2.68 (m, 2H), 3.27 (s, 3H), 3.62-3.73 (m, 3H), 3.87 (s,3H), 4.51-4.53 (m, 1H), 6.94 (s, 1H), 6.99 (d, J=8.8 Hz, 2H), 7.14 (d,J=8.4 Hz, 2H).

Preparation 67E: tert-butyl1-(5-(4-cyano-3-fluorophenyl)-6-(4-methoxyphenyl)-1-methyl-2-oxo-1,2-dihydropyridin-3-yl)piperidin-4-ylcarbamate

A mixture oftert-butyl-1-(5-bromo-6-(4-methoxyphenyl)-1-methyl-2-oxo-1,2-dihydropyridin-3-yl)piperidin-4-ylcarbamate(0.65 g, 1.32 mmol), tert-butyl piperidin-4-ylcarbamate (436 mg, 2.64mmol), Pd(dppf)₂Cl₂ (95 mg, 0.13 mmol) and Na₂CO₃ (420 mg, 3.96 mmol) inDMF/H₂O (20 mL/3 mL) was flushed with nitrogen and stirred at 100° C.for 2 h. The reaction mixture was cooled to RT and the insolublematerials filtered. The solvent was concentrated in vacuo and theresidue was purified by silica chromatography (1:1, EA:PE) to give 0.20g (28%) of the title compound. [M+H] Calc'd for C₃₀H₃₃FN₄O₄, 533; Found,533.

Example 67:4-(5-(4-aminopiperidin-1-yl)-2-(4-methoxyphenyl)-1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorobenzonitrile

To a solution of tert-butyl1-(5-(4-cyano-3-fluorophenyl)-6-(4-methoxyphenyl)-1-methyl-2-oxo-1,2-dihydropyridin-3-yl)piperidin-4-ylcarbamate(0.2 g, 0.38 mmol) in EA (10 mL) was added a solution of 4N HCl in EA (5mL), and the mixture was stirred at RT for 2 h. The solvent wasconcentrated in vacuo and the residue was purified by preparative HPLCto give 80 mg (45%) of the title compound as the HCl salt. ¹H NMR (400MHz, CDCl₃): δ 2.14-2.23 (m, 2H), 2.31-2.35 (m, 2H), 3.41 (s, 3H),3.53-3.65 (m, 3H), 3.80 (s, 3H), 3.88-3.91 (m, 2H), 6.95 (d, J=8.4 Hz,2H), 7.04-7.12 (m, 2H), 7.16 (d, J=8.8 Hz, 2H), 7.53-7.57 (m, 1H), 7.85(s, 1H). [M+H] Calc'd for C₂₅H₂₅FN₄O₂, 433; Found, 433.

Example 68:5-(5-(4-aminopiperidin-1-yl)-2-(4-methoxyphenyl)-1-methyl-6-oxo-1,6-dihydropyridin-3-yl)picolinonitrile

The title compound was prepared as the HCl salt in 13% overall yieldaccording to the general procedure for the preparation of Example 67. ¹HNMR (400 MHz, MeOD-d₄): δ 2.10-2.15 (m, 2H), 2.28-2.31 (m, 2H),3.33-3.34 (m, 1H), 3.48-3.51 (m, 5H), 3.83 (s, 3H), 3.88-3.91 (m, 2H),6.98 (d, J=8.8 Hz, 2H), 7.19 (d, J=8.8 Hz, 2H), 7.53-7.57 (m, 1H), 7.64(s, 1H), 7.72-7.76 (m, 2H), 7.38 (t, J=0.8 Hz, 1H). [M+H] Calc'd forC₂₄H₂₄N₅O₂, 416; Found, 416.

Example 69:(S)-5-(2-(4-methoxyphenyl)-1-methyl-6-oxo-5-(pyrrolidin-3-ylmethylamino)-1,6-dihydropyridin-3-yl)picolinonitrile

The title compound was prepared as the HCl salt in 22% overall yieldaccording to the general procedure for the preparation of Example 67. ¹HNMR (400 MHz, MeOD-d₄): δ 1.89-1.94 (m, 1H), 2.32-2.36 (m, 1H),2.87-2.91 (m, 1H), 3.14-3.19 (m, 1H), 3.34-3.35 (m, 1H), 3.43 (s, 3H),3.45-3.50 (m, 3H), 3.56-3.61 (m, 1H), 3.82 (s, 3H), 6.96 (d, J=8.4 Hz,2H), 7.18-7.22 (m, 3H), 7.72-7.78 (m, 2H), 7.54 (t, J=7.2 Hz, 1H). [M+H]Calc'd for C₂₄H₂₅N₅O₂, 416; Found, 416.

Example 70:(S)-2-fluoro-4-(2-(4-methoxyphenyl)-1-methyl-6-oxo-5-(pyrrolidin-3-ylmethylamino)-1,6-dihydropyridin-3-yl)benzonitrile

The title compound was prepared as the HCl salt in 44% overall yieldaccording to the general procedure for the preparation of Example 67. ¹HNMR (400 MHz, MeOD-d₄): δ 1.88-1.94 (m, 1H), 2.32-2.36 (m, 1H),2.86-2.90 (m, 1H), 3.14-3.19 (m, 1H), 3.35-3.38 (m, 1H), 3.42 (s, 3H),3.44-3.51 (m, 3H), 3.56-3.61 (m, 1H), 3.83 (s, 3H), 6.96 (d, J=8.8 Hz,2H), 7.07-7.13 (m, 2H), 7.17-7.19 (m, 3H), 8.38 (s, 1H). [M+H] Calc'dfor C₂₅H₂₅FN₄O₂, 433; Found, 433.

Example 71:(S)-5-(2-(4-ethylphenyl)-1-methyl-6-oxo-5-(pyrrolidin-3-ylmethylamino)-1,6-dihydropyridin-3-yl)picolinonitrile

The title compound was prepared as the HCl salt in 15% overall yieldaccording to the general procedure for the preparation of Example 67. ¹HNMR (400 MHz, MeOD-d₄): d 1.23 (t, J=7.6 Hz, 3H), 1.86-1.91 (m, 1H),2.28-2.32 (m, 1H), 2.65 (q, J=7.6 Hz, 2H), 2.83-2.87 (m, 1H), 3.10-3.15(m, 1H), 3.37-3.47 (m, 7H), 3.51-3.56 (m, 1H), 6.81 (s, 1H), 7.15 (d,J=7.6 Hz, 2H), 7.25 (d, J=8.0 Hz, 2H), 7.68-7.74 (m, 2H), 8.36 (s, 1H).[M+H] Calc'd for C₂₅H₂₇N₅O, 414; Found, 414.

Example 72:(S)-4-(2-(4-ethylphenyl)-1-methyl-6-oxo-5-(pyrrolidin-3-ylmethylamino)-1,6-dihydropyridin-3-yl)-2-fluorobenzonitrile

The title compound was prepared as the HCl salt in 27% overall yieldaccording to the general procedure for the preparation of Example 67. ¹HNMR (400 MHz, MeOD-d₄): δ 1.19 (t, J=7.6 Hz, 3H), 1.84-1.88 (m, 1H),2.26-2.31 (m, 1H), 2.62 (q, J=7.6 Hz, 2H), 2.81-2.85 (m, 1H), 3.09-3.13(m, 1H), 3.34-3.46 (m, 7H), 3.51-3.55 (m, 1H), 7.00-7.06 (m, 3H), 7.12(d, J=8.0 Hz, 2H), 7.22 (d, J=8.0 Hz, 2H), 7.48 (t, J=7.6 Hz, 1H). [M+H]Calc'd for C₂₆H₂₇FN₄O, 431; Found, 431.

Example 73:(S)-5-(2-(4-cyclopropylphenyl)-1-methyl-6-oxo-5-(pyrrolidin-3-ylmethylamino)-1,6-dihydropyridin-3-yl)picolinonitrile

The title compound was prepared as the HCl salt in 13% overall yieldaccording to the general procedure for the preparation of Example 67.¹HNMR (400 MHz, MeOD-d₄): δ 0.70-0.73 (m, 2H), 1.02-1.05 (m, 2H),1.89-1.94 (m, 2H), 2.27-2.32 (m, 1H), 2.84-2.88 (m, 1H), 3.11-3.16 (m,1H), 3.37-3.49 (m, 7H), 3.51-3.56 (m, 1H), 6.85 (s, 1H), 7.11 (s, 4H),7.69-7.75 (m, 2H), 8.36 (s, 1H). [M+H] Calc'd for C₂₆H₂₇FN₄O, 426;Found, 426.

Example 74:(S)-4-(2-(4-cyclopropylphenyl)-1-methyl-6-oxo-5-(pyrrolidin-3-ylmethylamino)-1,6-dihydropyridin-3-yl)-2-fluorobenzonitrile

The title compound was prepared as the HCl salt in 29% overall yieldaccording to the general procedure for the preparation of Example 67. ¹HNMR (400 MHz, MeOD-d₄): δ 0.70-0.74 (m, 2H), 1.01-1.06 (m, 2H),1.87-1.96 (m, 2H), 2.31-2.34 (m, 1H), 2.85-2.88 (m, 1H), 3.12-3.17 (m,1H), 3.35-3.49 (m, 7H), 3.54-3.59 (m, 1H), 6.93-6.94 (m, 1H), 7.06-7.12(m, 6H), 7.52 (t, J=7.2 Hz, 1H). [M+H] Calc'd for C₂₇H₂₇N₅O, 443; Found,443.

Example 75:4-[5-(4-aminopiperidin-1-yl)-1-methyl-2-(4-methylphenyl)-6-oxopyridin-3-yl]benzonitrile

The title compound was prepared as the HCl salt in 16% overall yieldaccording to the general procedure for the preparation of Example 67. ¹HNMR (400 MHz, MeOD-d₄): δ 2.33-2.37 (m, 2H), 2.44-2.47 (m, 2H), 3.44 (s,3H), 3.69-3.70 (m, 1H), 3.96-4.02 (m, 4H), 7.16 (d, J=8.0 Hz, 2H), 7.25(d, J=7.6 Hz, 2H), 7.30 (d, J=8.4 Hz, 2H), 7.55 (d, J=8.0 Hz, 2H), 8.17(s, 1H). [M+H] Calc'd for C₂₅H₂₆N₄O, 399; Found, 399.

Example 76:4-[5-(4-aminopiperidin-1-yl)-1-methyl-2-(4-methylphenyl)-6-oxopyridin-3-yl]-2-fluorobenzonitrile

The title compound was prepared as the HCl salt in 11% overall yieldaccording to the general procedure for the preparation of Example 67. ¹HNMR (400 MHz, MeOD-d₄): δ 2.25-2.28 (m, 2H), 2.37-2.40 (m, 5H), 3.44 (s,3H), 3.61-3.63 (m, 1H), 3.78-3.81 (m, 2H), 3.93-3.96 (m, 2H), 7.09-7.19(m, 4H), 7.28 (d, J=7.6 Hz, 2H), 7.56 (t, J=7.2 Hz, 1H), 8.17 (s, 1H).[M+H] Calc'd for C₂₅H₂₅FN₄O, 417; Found, 417.

Example 77:4-[5-(4-aminopiperidin-1-yl)-2-(4-methoxyphenyl)-1-methyl-6-oxopyridin-3-yl]benzonitrile

The title compound was prepared as the HCl salt in 13% overall yieldaccording to the general procedure for the preparation of Example 67. ¹HNMR (400 MHz, MeOD-d₄): δ 2.19-2.23 (m, 2H), 2.30-2.33 (m, 2H), 3.34 (s,3H), 3.51-3.58 (m, 1H), 3.70 (s, 3H), 3.83-3.88 (m, 4H), 6.84 (d, J=8.8Hz, 2H), 7.06 (d, J=8.4 Hz, 2H), 7.17 (d, J=8.0 Hz, 2H), 7.44 (d, J=8.4Hz, 2H), 8.03 (s, 1H). [M+H] Calc'd for C₂₅H₂₆N₄O₂, 415; Found, 415.

Example 78:4-{2-(4-methoxyphenyl)-1-methyl-5-[4-(methylamino)piperidyl]-6-oxo-3-hydropyridyl}benzenecarbonitrile

The title compound was prepared as the HCl salt in 14% overall yieldaccording to the general procedure for the preparation of Example 67. ¹HNMR (400 MHz, CD₃OD): δ 2.14-2.18 (m, 2H), 2.41-2.44 (m, 2H), 2.82 (s,3H), 3.45 (s, 3H), 3.47-3.55 (m, 3H), 3.82 (s, 3H), 3.90-3.96 (m, 2H),6.96 (d, J=8.8 Hz, 2H), 7.17 (d, J=8.4 Hz, 2H), 7.28 (d, J=8.0 Hz, 2H),7.57 (d, J=8.0 Hz, 2H), 7.76 (s, 1H). [M+H] Calc'd for C₂₆H₂₈N₄O₂, 429;Found, 429.

Example 79:2-fluoro-4-{2-(4-methoxyphenyl)-1-methyl-5-[4-(methylamino)piperidyl]-6-oxo(3-hydropyridyl)}benzenecarbonitrile

The title compound was prepared as the HCl salt in 78% overall yieldaccording to the general procedure for the preparation of Example 67. ¹HNMR (400 MHz, CD₃OD): δ 2.12-2.16 (m, 2H), 2.40-2.43 (m, 2H), 2.82 (s,3H), 3.45 (s, 3H), 3.46-3.52 (m, 3H), 3.84 (s, 3H), 3.90-3.96 (m, 2H),6.99 (d, J=8.8 Hz, 2H), 7.08 (d, J=8.0 Hz, 1H), 7.13 (d, J=10.8 Hz, 1H),7.20 (d, J=8.8 Hz, 2H), 7.58 (t, J=7.2 Hz, 1H), 7.72 (s, 1H). [M+H]Calc'd for C₂₆H₂₇FN₄O₂, 447; Found, 447.

Example 80:4-{1-methyl-5-[4-(methylamino)piperidyl]-2-(4-methylphenyl)-6-oxo-3-hydropyridyl}benzenecarbonitrile

The title compound was prepared as the HCl salt in 18% overall yieldaccording to the general procedure for the preparation of Example 67. ¹HNMR (300 MHz, CD₃OD): δ 2.14-2.24 (m, 2H), 2.36 (s, 3H), 2.42-2.46 (m,2H), 2.80 (s, 3H), 3.42 (s, 3H), 3.57-3.58 (m, 3H), 3.92-3.96 (m, 2H),7.14 (d, J=8.1 Hz, 2H), 7.22-7.29 (m, 4H), 7.53 (d, J=9.6 Hz, 2H), 7.82(s, 1H). [M+H] Calc'd for C₂₆H₂₈N₄O, 413; Found, 413.

Example 81:4-[5-(4-aminopiperidin-1-yl)-1-(cyclopropylmethyl)-2-(4-methoxyphenyl)-6-oxopyridin-3-yl]benzonitrile

The title compound was prepared as the HCl salt in 21% overall yieldaccording to the general procedure for the preparation of Example 67. ¹HNMR (300 MHz, CD₃OD): δ 0.14-0.18 (m, 2H), 0.37-0.41 (m, 2H), 0.98-1.02(m, 1H), 2.16-2.23 (m, 2H), 2.32-2.35 (m, 2H), 3.51-3.59 (m, 3H), 3.79(s, 3H), 3.88-3.92 (m, 2H), 3.99-4.01 (m, 2H), 6.92 (d, J=8.6 Hz, 2H),7.18 (d, J=8.4 Hz, 2H), 7.26 (d, J=8.0 Hz, 2H), 7.54 (d, J=8.0 Hz, 2H),7.77 (s, 1H). [M+H] Calc'd for C₂₈H₃₀N₄O₂, 455; Found, 455.

Example 82:4-[5-(4-aminopiperidin-1-yl)-1-ethyl-2-(4-methoxyphenyl)-6-oxopyridin-3-yl]benzonitrile

The title compound was prepared as the HCl salt in 21% overall yieldaccording to the general procedure for the preparation of Example 67. ¹HNMR (300 MHz, CD₃OD): δ 1.20 (t, J=7.2 Hz, 3H), 2.23-2.29 (m, 2H),2.37-2.40 (m, 2H), 3.57-3.63 (m, 3H), 3.71-3.76 (m, 2H), 3.82 (s, 3H),3.92-3.95 (m, 2H), 4.06 (t, J=7.2 Hz, 2H), 6.96 (d, J=8.0 Hz, 2H), 7.22(d, J=8.8 Hz, 2H), 7.29 (d, J=8.8 Hz, 2H), 7.56 (d, J=8.0 Hz, 2H), 7.92(s, 1H). [M+H] Calc'd for C₂₆H₂₈N₄O₂, 429; Found, 429.

Example 83:4-[5-(4-aminopiperidin-1-yl)-2-(3-fluoro-4-methoxyphenyl)-1-methyl-6-oxopyridin-3-yl]benzonitrile

The title compound was prepared as the HCl salt in 10% overall yieldaccording to the general procedure for the preparation of Example 67. ¹HNMR (400 MHz, MeOD-d₄): δ 2.08-2.11 (m, 2H), 2.22-2.25 (m, 2H), 3.39 (s,3H), 3.48-3.52 (m, 3H), 3.77-3.85 (m, 5H), 6.86 (d, J=8.4 Hz, 1H), 6.98(t, J=8.8 Hz, 2H), 7.18 (d, J=8.0 Hz, 2H), 7.47 (d, J=8.0 Hz, 2H),7.70-7.75 (m, 1H). [M+H] Calc'd for C₂₅H₂₅FN₄O₂, 433; Found, 433.

Example 84:4-[5-(4-aminopiperidin-1-yl)-2-(3-fluoro-4-methoxyphenyl)-1-methyl-6-oxopyridin-3-yl]-2-fluorobenzonitrile

The title compound was prepared as the HCl salt in 9% overall yieldaccording to the general procedure for the preparation of Example 67. ¹HNMR (400 MHz, MeOD-d₄): δ 2.16-2.19 (m, 2H), 2.32-2.35 (m, 2H), 3.45 (s,3H), 3.50-3.58 (m, 3H), 3.91-3.95 (m, 5H), 7.01 (d, J=8.4 Hz, 1H),7.10-7.20 (m, 4H), 7.59 (t, J=7.2 Hz, 1H), 7.77 (s, 1H). [M+H] Calc'dfor C₂₅H₂₄F₂N₄O₂, 451; Found, 451.

Example 85:4-[5-(4-aminopiperidin-1-yl)-1-(cyclopropylmethyl)-2-(4-methoxyphenyl)-6-oxopyridin-3-yl]-2-fluorobenzonitrile

The title compound was prepared as the HCl salt in 21% overall yieldaccording to the general procedure for the preparation of Example 67. ¹HNMR (300 MHz, CD₃OD): δ 0.14-0.17 (m, 2H), 0.37-0.41 (m, 2H), 0.96-1.02(m, 1H), 2.14-2.24 (m, 2H), 2.33-2.36 (m, 2H), 3.53-3.64 (m, 3H), 3.81(s, 3H), 3.88-3.92 (m, 2H), 3.99-4.01 (m, 2H), 6.96 (d, J=8.8 Hz, 2H),7.06-7.14 (m, 2H), 7.21 (d, J=8.4 Hz, 2H), 7.53-7.57 (m, 1H), 7.83 (s,1H). [M+H] Calc'd for C₂₈H₂₉FN₄O₂, 473; Found, 473.

Example 86:2-fluoro-4-[2-(3-fluoro-4-methoxyphenyl)-1-methyl-5-[4-(methylamino)piperidin-1-yl]-6-oxopyridin-3-yl]benzonitrile

The title compound was prepared as the HCl salt in 7% overall yieldaccording to the general procedure for the preparation of Example 67.¹HNMR (400 MHz, MeOD-d₄): δ 2.23-2.23 (m, 2H), 2.47-2.50 (m, 2H), 2.82(s, 3H), 3.46 (s, 3H), 3.52-3.57 (m, 1H), 3.71-3.77 (m, 2H), 3.92-3.98(m, 5H), 7.02 (d, J=8.4 Hz, 1H), 7.10-7.22 (m, 4H), 7.60 (t, J=7.2 Hz,1H), 7.96 (s, 1H). [M+H] Calc'd for C₂₆H₂₆F₂N₄O₂, 465; Found, 465.

Example 87:4-[5-(4-aminopiperidin-1-yl)-1-ethyl-2-(4-methoxyphenyl)-6-oxopyridin-3-yl]-2-fluorobenzonitrile

The title compound was prepared as the HCl salt in 25% overall yieldaccording to the general procedure for the preparation of Example 67. ¹HNMR (300 MHz, CD₃OD): δ 1.20 (t, J=7.2 Hz, 3H), 2.21-2.24 (m, 2H),2.36-2.39 (m, 2H), 3.68-3.75 (m, 3H), 3.84 (s, 3H), 3.92-3.95 (m, 2H),4.06 (q, J=7.2 Hz, 2H), 7.00 (d, J=8.8 Hz, 2H), 7.09-7.17 (m, 2H), 7.25(d, J=8.4 Hz, 2H), 7.58 (t, J=7.6 Hz, 1H), 7.89 (s, 1H). [M+H] Calc'dfor C₂₈H₂₉FN₄O₂, 473; Found, 473.

Example 88:2-[3-(4-aminopiperidyl)-5-(4-cyano-3-fluorophenyl)-6-(4-methoxyphenyl)-2-oxohydropyrazinyl]acetamide

The title compound was prepared as the HCl salt in 13% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.74-1.78 (m, 2H), 2.07-2.10 (m, 2H), 2.99-3.06(m, 2H), 3.34-3.41 (m, 1H), 3.83 (s, 3H), 4.46 (s, 2H), 4.86-4.90 (m,2H), 6.99 (d, J=8.0 Hz, 2H), 7.10 (dd, J=1.6 Hz, 8.0 Hz, 1H), 7.16-7.25(m, 3H), 7.44-7.48 (m, 1H). [M+H] Calc'd for C₂₅H₂₅FN₆O₃, 476; Found,477.

Example 89:3-[3-(4-aminopiperidyl)-5-(4-cyano-3-fluorophenyl)-6-(4-methoxyphenyl)-2-oxohydropyrazinyl]propanamide

The title compound was prepared as the HCl salt in 16% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.72-1.78 (m, 2H), 2.03-2.05 (m, 2H), 2.43 (t,J=8.0 Hz, 2H), 3.01-3.07 (m, 2H), 3.37-3.41 (m, 1H), 3.73 (s, 3H), 3.96(t, J=8.0 Hz, 2H), 4.76-4.80 (m, 2H), 6.90 (d, J=8.8 Hz, 2H), 7.04-7.17(m, 4H), 7.46 (t, J=7.2 Hz, 1H). [M+H] Calc'd for C₂₆H₂₇FN₆O₃, 490;Found, 491.

Example 90:4-[3-(4-aminopiperidyl)-5-(4-cyano-3-fluorophenyl)-6-(4-methoxyphenyl)-2-oxohydropyrazinyl]butanamide

The title compound was prepared as the free base in 11% overall yieldaccording to the general procedure for the preparation of Example 16. ¹HNMR (400 MHz, CD₃OD): δ 1.52-1.55 (m, 2H), 1.81-1.86 (m, 2H), 1.96-2.05(m, 2H), 2.06 (t, J=7.2 Hz, 2H), 2.96-3.02 (m, 3H), 3.85 (s, 3H),3.85-3.88 (m, 2H), 4.76-4.80 (m, 2H), 7.00 (d, J=8.8 Hz, 2H), 7.09-7.26(m, 4H), 7.44 (t, J=8.0 Hz, 1H). [M+H] Calc'd for C₂₇H₂₉FN₆O₃, 504;Found, 505.

Example 91:4-{5-(4-aminopiperidyl)-2-[6-(dimethylamino)(3-pyridyl)]-1-methyl-6-oxo(3-hydropyridyl)}-2-fluorobenzenecarbonitrile

The title compound was prepared as the free base in 14% overall yieldaccording to the general procedure for the preparation of Example 67. ¹HNMR (400 MHz, CD₃OD): δ 2.47-2.30 (m, 2H), 2.39-2.42 (m, 2H), 3.35 (s,6H), 3.53 (s, 3H), 3.59-3.71 (m, 3H), 3.95-3.98 (m, 2H), 7.22 (d, J=8.0Hz, 1H), 7.38 (t, J=10.4 Hz, 2H), 7.71 (t, J=7.2 Hz, 1H), 7.89 (s, 1H),7.92 (s, 1H), 8.03 (d, J=9.2 Hz, 1H). [M+H] Calc'd for C₂₅H₂₇FN₆O, 446;Found, 447.

Example 92:4-{2-[6-(dimethylamino)(3-pyridyl)]-1-methyl-5-[4-(methylamino)piperidyl]-6-oxo-3-hydropyridyl}benzenecarbonitrile

The title compound was prepared as the free base in 16% overall yieldaccording to the general procedure for the preparation of Example 67. ¹HNMR (400 MHz, CD3OD): δ 2.18-2.24 (m, 2H), 2.40-2.47 (m, 2H), 2.83 (s,3H), 3.35 (s, 6H), 3.51-3.54 (m, 3H), 3.54 (s, 3H), 3.98-4.01 (m, 2H),7.35 (d, J=8.8 Hz, 1H), 7.38 (d, J=8.0 Hz, 2H), 7.71 (d, J=8.0 Hz, 2H),7.76 (s, 1H), 7.86 (s, 1H), 8.02 (d, J=8.2 Hz, 1H). [M+H] Calc'd forC₂₆H₃₀N₆O, 442; Found, 443.

Chemical MS Synthesis Structure (ESI) Example (prepared by procedure ofcited Example) m/z 93

418 94

417 95

405 96

443 97

425

II. Biological Evaluation Example 1a: In Vitro Enzyme InhibitionAssay—LSD-1

This assay determines the ability of a test compound to inhibit LSD1demethylase activity. E. coli expressed full-length human LSD1(Accession number 060341) was purchased from Active Motif (Cat#31334).

The enzymatic assay of LSD1 activity is based on TimeResolved-Fluorescence Resonance Energy Transfer (TR-FRET) detection. Theinhibitory properties of compounds to LSD1 were determined in 384-wellplate format under the following reaction conditions: 0.1-0.5 nM LSD1,50 nM H3K4me1-biotin labeled peptide (Anaspec cat #64355), 2 μM FAD inassay buffer of 50 mM HEPES, pH7.3, 10 mM NaCl, 0.005% Brij35, 0.5 mMTCEP, 0.2 mg/ml BSA. Reaction product was determined quantitatively byTR-FRET after the addition of detection reagent PhycolinkStreptavidin-allophycocyanin (Prozyme) and Europium-anti-unmodifiedhistone H3 lysine 4 (H3K4) antibody (PerkinElmer) in the presence ofLSD1 inhibitor such as 1.8 mM of Tranylcypromine hydrochloride (2-PCPA)in LANCE detection buffer (PerkinElmer) to final concentration of 12.5nM and 0.25 nM respectively.

The assay reaction was performed according to the following procedure: 2μL of the mixture of 150 nM H3K4me1-biotin labeled peptide with 2 μL of11-point serial diluted test compound in 3% DMSO were added to each wellof plate, followed by the addition of 2 μL of 0.3 nM LSD1 and 6 μM ofFAD to initiate the reaction. The reaction mixture was then incubated atroom temperature for one hour, and terminated by the addition of 6 μL of1.8 mM 2-PCPA in LANCE detection buffer containing 25 nM PhycolinkStreptavidin-allophycocyanin and 0.5 nM Europium-anti-unmodified H3K4antibody. Enzymatic reaction is terminated within 15 minutes if 0.5 LSD1enzyme is used in the plate. Plates were read by EnVision MultilabelReader in TR-FRET mode (excitation at 320 nm, emission at 615 nm and 665nm) after 1 hour incubation at room temperature. A ratio was calculated(665/615) for each well and fitted to determine inhibition constant(IC₅₀).

The ability of the compounds disclosed herein to inhibit LSD1 activitywas quantified and the respective IC₅₀ value was determined. Table 3provides the IC₅₀ values of various substituted heterocyclic compoundsdisclosed herein.

TABLE 3 Chemical LSD1 Synthesis IC₅₀ Example Name (μM) 14-[6-(4-aminopiperidin-1-yl)-4-benzyl-5-oxo-4,5- Adihydropyrazin-2-yl]benzonitrile 2 4-[6-(4-aminopiperidin-1-yl)-4-[(4- Amethylphenyl)methyl]-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 34-[6-(4-aminopiperidin-1-yl)-4-[(2- Afluorophenyl)methyl]-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 44-[6-(4-aminopiperidin-1-yl)-4-(3-chloro-benzyl)-5- Aoxo-4,5-dihydro-pyrazin-2-yl]-benzonitrile 54-[6-(4-amino-piperidin-1-yl)-4-methyl-5-oxo-4,5- Bdihydro-pyrazin-2-yl]-2-fluoro-benzonitrile 64-[6-(4-Amino-piperidin-1-yl)-4-ethyl-5-oxo-4,5- Bdihydro-pyrazin-2-yl]-2-fluoro-benzonitrile 74-[6-(4-amino-piperidin-1-yl)-4-cyclopropylmethyl-5- Aoxo-4,5-dihydro-pyrazin-2-yl]-2-fluoro-benzonitrile 84-[6-(4-aminopiperidin-1-yl)-4-[(3- Afluorophenyl)methyl]-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 94-[6-(4-aminopiperidin-1-yl)-4-[(4- Afluorophenyl)methyl]-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 104-[6-(4-aminopiperidin-1-yl)-4-[(3- Amethoxyphenyl)methyl]-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 114-[6-(4-aminopiperidin-1-yl)-4-[(4- Amethoxyphenyl)methyl]-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 124-[6-(4-aminopiperidin-1-yl)-5-oxo-4-[(1R)-1- Aphenylethyl]-4,5-dihydropyrazin-2-yl]benzonitrile 134-[6-(4-aminopiperidin-1-yl)-5-oxo-4-[(1S)-1- Aphenylethyl]-4,5-dihydropyrazin-2-yl]benzonitrile 144-[6-(4-amino-piperidin-1-yl)-5-oxo-4-(tetrahydro- Afuran-3-ylmethyl)-4,5-dihydro-pyrazin-2-yl]-2-fluoro- benzonitrile 154-[6-(4-amino-piperidin-1-yl)-5-oxo-4-(tetrahydro- Apyran-4-ylmethyl)-4,5-dihydro-pyrazin-2-yl]-2-fluoro- benzonitrile 164-[6-(4-aminopiperidin-1-yl)-4-methyl-3-(4- Amethylphenyl)-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 174-[4-methyl-3-(4-methylphenyl)-5-oxo-6-{[(3S)- Apyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin-2- yl]benzonitrile 184-[4-methyl-3-(4-methylphenyl)-5-oxo-6-{[(3R)- Apyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin-2- yl]benzonitrile 195-[6-(4-aminopiperidin-1-yl)-4-methyl-3-(4-methyl- Aphenyl)-5-oxo-4,5-dihydropyrazin-2-yl]pyridine- 2-carbonitrile 204-{4-methyl-6-[4-(methylamino)piperidin-1-yl]- A3-(4-methylphenyl)-5-oxo-4,5-dihydropyrazin-2- yl}benzonitrile 214-[6-(4-amino-4-methylpiperidin-1-yl)-4-methyl-3- A(4-methylphenyl)-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 224-[6-(3-aminopiperidin-1-yl)-4-methyl-3-(4- Amethylphenyl)-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 234-[4-methyl-3-(4-methylphenyl)-6-{octahydro- A1H-pyrrolo[3,4-c]pyridin-5-yl}-5-oxo-4,5-dihydropyrazin-2-yl]benzonitrile 244-(6-{2,8-diazaspiro[4.5]decan-8-yl}-4-methyl- A3-(4-methylphenyl)-5-oxo-4,5-dihydropyrazin-2- yl)benzonitrile 254-(6-{decahydropyrrolo[3,4-d]azepin-6-yl}- A4-methyl-3-(4-methylphenyl)-5-oxo-4,5-dihydro- pyrazin-2-yl)benzonitrile26 4-[6-(4-aminopiperidin-1-yl)-4-ethyl-3-(4- Amethylphenyl)-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 274-[6-(4-aminopiperidin-1-yl)-3-(4-ethylphenyl)-4- Amethyl-5-oxo-4,5-dihydropyrazin-2-yl]benzonitrile 284-[3-(4-ethylphenyl)-4-methyl-5-oxo-6-{[(3S)- Apyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin- 2-yl]benzonitrile 294-[3-(4-ethylphenyl)-4-methyl-5-oxo-6-{[(3R)- Apyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin- 2-yl]benzonitrile 304-[6-(4-aminopiperidin-1-yl)-3-(4-methoxyphenyl)-4- Amethyl-5-oxo-4,5-dihydropyrazin-2-yl]benzonitrile 314-[3-(4-methoxyphenyl)-4-methyl-5-oxo-6- A{[(3S)-pyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin-2-yl]benzonitrile 324-[3-(4-methoxyphenyl)-4-methyl-5-oxo-6- A{[(3R)-pyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin-2-yl]benzonitrile 334-[6-(4-aminopiperidin-1-yl)-3-(4-cyclopropyl- Aphenyl)-4-methyl-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 344-[3-(4-cyclopropylphenyl)-4-methyl-5-oxo-6- A{[(3S)-pyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin-2-yl]benzonitrile 354-[3-(4-cyclopropylphenyl)-4-methyl-5-oxo-6- A{[(3R)-pyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin-2-yl]benzonitrile 364-[6-(4-amino-piperidin-1-yl)-4-(2-methoxy- Aethyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2- yl]-benzonitrile 374-[6-(4-amino-piperidin-1-yl)-4-(2-hydroxy- Aethyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2- yl]-benzonitrile 384-[6-(4-amino-piperidin-1-yl)-3-(4-cyclopropyl- Aphenyl)-4-(2-hydroxy-ethyl)-5-oxo-4,5-dihydro-pyrazin-2-yl]-benzonitrile 394-[6-(4-amino-piperidin-1-yl)-4-(3-methoxy- Apropyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2- yl]-benzonitrile 404-[6-(4-amino-piperidin-1-yl)-4-(3-hydroxy- Apropyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2- yl]-benzonitrile 414-[6-(4-amino-piperidin-1-yl)-4-(2-methoxy- Aethyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2- yl]-2-fluoro-benzonitrile42 4-[6-(4-amino-piperidin-1-yl)-4-(2-hydroxy- Aethyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2- yl]-2-fluoro-benzonitrile43 4-(6-{[((3S)-pyrrolidin-3-yl)methyl]amino}- A4-(2-methoxyethyl)-3-(4-methylphenyl)-5-oxo(4-hydropyrazin-2-yl))-2-fluorobenzenecarbonitrile 444-(6-{[((3S)-pyrrolidin-3-yl)methyl]amino}- A4-(2-hydroxyethyl)-3-(4-methylphenyl)-5-oxo(4-hydropyrazin-2-yl))-2-fluorobenzenecarbonitrile 454-[6-(4-aminopiperidyl)-4-(3-methoxypropyl)- A3-(4-methylphenyl)-5-oxo(4-hydropyrazin-2-yl)]-2-fluorobenzenecarbonitrile 464-[6-(4-aminopiperidyl)-4-(3-hydroxypropyl)- A3-(4-methylphenyl)-5-oxo(4-hydropyrazin-2-yl)]-2-fluorobenzenecarbonitrile 474-[6-(4-amino-piperidin-1-yl)-4-methyl-5- Aoxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]-2- fluoro-benzonitrile 484-[6-(4-amino-piperidin-1-yl)-4-methyl-5- Aoxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]-3- fluoro-benzonitrile 494-[6-(4-amino-piperidin-1-yl)-4-methyl-3- A(6-methyl-pyridin-3-yl)-5-oxo-4,5-dihydro- pyrazin-2-yl]-benzonitrile 504-[6-(4-amino-piperidin-1-yl)-3-(4-cyclopropyl- Aphenyl)-4-methyl-5-oxo-4,5-dihydro-pyrazin- 2-yl]-2-fluoro-benzonitrile51 5-[6-(4-amino-piperidin-1-yl)-3-(4-methoxy- Aphenyl)-4-methyl-5-oxo-4,5-dihydro-pyrazin-2-yl]-pyridine-2-carbonitrile 524-[6-(4-amino-piperidin-1-yl)-3-(4-methoxy- Aphenyl)-4-methyl-5-oxo-4,5-dihydro-pyrazin-2- yl]-2-fluoro-benzonitrile53 5-[6-(4-amino-piperidin-1-yl)-3-(4-cyclopropyl- Aphenyl)-4-methyl-5-oxo-4,5-dihydro-pyrazin-2-yl]-pyridine-2-carbonitrile 54 5-{3-(4-methoxy-phenyl)-4-methyl-5-oxo-6-A [((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro-pyrazin-2-yl}-pyridine-2-carbonitrile 552-fluoro-4-(3-(4-methoxy-phenyl)-4-methyl-5- Aoxo-6-[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro-pyrazin-2-yl}-benzonitrile 565-{3-(4-ethyl-phenyl)-4-methyl-5-oxo-6- A[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro-pyrazin-2-yl}-pyridine-2-carbonitrile 574-{3-(4-ethyl-phenyl)-4-methyl-5-oxo-6- A[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro-pyrazin-2-yl}-2-fluoro-benzonitrile 585-(3-(4-cyclopropyl-phenyl)-4-methyl-5-oxo-6- A[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro-pyrazin-2-yl}-pyridine-2-carbonitrile 594-{3-(4-cyclopropyl-phenyl)-4-methyl-5-oxo-6- A[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro-pyrazin-2-yl}-2-fluoro-benzonitrile 604-[6-(4-amino-piperidin-1-yl)-4-methyl-3- A(1-methyl-1H-benzoimidazol-5-yl)-5-oxo-4,5-dihydro-pyrazin-2-yl]-2-fluoro-benzonitrile 614-{3-(4-cyclopropylmethoxy-phenyl)-4- Amethyl-5-oxo-6-[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro-pyrazin-2-yl}-2-fluoro- benzonitrile 624-{3-(4-ethoxy-phenyl)-4-methyl-5-oxo-6- A[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro-pyrazin-2-yl}-2-fluoro-benzonitrile 632-fluoro-4-[4-methyl-5-oxo-6-[((3S)- Apyrrolidin-3-ylmethyl)-amino]-3-(4-trifluoro-methoxy-phenyl)-4,5-dihydro-pyrazin-2-yl]- benzonitrile 642-Fluoro-4-[4-(3-hydroxy-propyl)-6-(4-methyl- Aamino-piperidin-1-yl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]-benzonitrile 652-Fluoro-4-[4-(2-hydroxy-ethyl)-6-(4-methylamino- Apiperidin-1-yl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin- 2-yl]-benzonitrile66 4-[6-(4-Amino-piperidin-1-yl)-4-methyl-3-(6- Amethyl-pyridin-3-yl)-5-oxo-4,5-dihydro-pyrazin-2-yl]-2-fluoro-benzonitrile 674-(5-(4-aminopiperidin-1-yl)-2-(4-methoxyphenyl)- A1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2- fluorobenzonitrile 685-(5-(4-aminopiperidin-1-yl)-2-(4-methoxyphenyl)- A1-methyl-6-oxo-1,6-dihydropyridin-3-yl)picolino- nitrile 69(S)-5-(2-(4-methoxyphenyl)-1-methyl-6-oxo-5- A(pyrrolidin-3-ylmethylamino)-1,6-dihydropyridin- 3-yl)picolinonitrile 70(S)-2-fluoro-4-(2-(4-methoxyphenyl)-1-methyl-6- Aoxo-5-(pyrrolidin-3-ylmethylamino)-1,6-dihydro-pyridin-3-yl)benzonitrile 71 (S)-5-(2-(4-ethylphenyl)-1-methyl-6-oxo-5-A (pyrrolidin-3-ylmethylamino)-1,6-dihydropyridin- 3-yl)picolinonitrile72 (S)-4-(2-(4-ethylphenyl)-1-methyl-6-oxo-5- A(pyrrolidin-3-ylmethylamino)-1,6-dihydropyridin-3-yl)-2-fluorobenzonitrile 73(S)-5-(2-(4-cyclopropylphenyl)-1-methyl-6-oxo- A5-(pyrrolidin-3-ylmethylamino)-1,6-dihydropyridin- 3-yl)picolinonitrile74 (S)-4-(2-(4-cyclopropylphenyl)-1-methyl-6-oxo- A5-(pyrrolidin-3-ylmethylamino)-1,6-dihydropyridin-3-yl)-2-fluorobenzonitrile 754-[5-(4-aminopiperidin-1-yl)-1-methyl-2-(4- Amethylphenyl)-6-oxopyridin-3-yl]benzonitrile 764-[5-(4-aminopiperidin-1-yl)-1-methyl-2-(4- Amethylphenyl)-6-oxopyridin-3-yl]-2-fluoro- benzonitrile 774-[5-(4-aminopiperidin-1-yl)-2-(4-methoxy- Aphenyl)-1-methyl-6-oxopyridin-3-yl]benzonitrile 784-{2-(4-methoxyphenyl)-1-methyl-5-[4- A (methylamino)piperidyl]-6-oxo-3-hydropyridyl}benzenecarbonitrile 792-fluoro-4-{2-(4-methoxyphenyl)-1-methyl-5- A[4-(methylamino)piperidyl]-6-oxo(3- hydropyridyl)}benzenecarbonitrile 804-{1-methyl-5-[4-(methylamino)piperidyl]- A 2-(4-methylphenyl)-6-oxo-3-hydropyridyl}benzenecarbonitrile 814-[5-(4-aminopiperidin-1-yl)-1-(cyclopropyl- Amethyl)-2-(4-methoxyphenyl)-6-oxopyridin- 3-yl]benzonitrile 824-[5-(4-aminopiperidin-1-yl)-1-ethyl-2-(4- Amethoxyphenyl)-6-oxopyridin-3-yl]benzonitrile 834-[5-(4-aminopiperidin-1-yl)-2-(3-fluoro-4- Amethoxyphenyl)-1-methyl-6-oxopyridin-3- yl]benzonitrile 844-[5-(4-aminopiperidin-1-yl)-2-(3-fluoro-4- Amethoxyphenyl)-1-methyl-6-oxopyridin-3-yl]-2- fluorobenzonitrile 854-[5-(4-aminopiperidin-1-yl)-1-(cyclopropyl- Amethyl)-2-(4-methoxyphenyl)-6-oxopyridin-3- yl]-2-fluorobenzonitrile 862-fluoro-4-[2-(3-fluoro-4-methoxyphenyl)-1- Amethyl-5-[4-(methylamino)piperidin-1-yl]- 6-oxopyridin-3-yl]benzonitrile87 4-[5-(4-aminopiperidin-1-yl)-1-ethyl-2-(4- Amethoxyphenyl)-6-oxopyridin-3-yl]-2-fluoro- benzonitrile 882-[3-(4-aminopiperidyl)-5-(4-cyano-3-fluoro- Aphenyl)-6-(4-methoxyphenyl)-2-oxohydro- pyrazinyl]acetamide 893-[3-(4-aminopiperidyl)-5-(4-cyano-3-fluoro- Aphenyl)-6-(4-methoxyphenyl)-2-oxohydro- pyrazinyl]propanamide 904-[3-(4-aminopiperidyl)-5-(4-cyano-3-fluoro- Aphenyl)-6-(4-methoxyphenyl)-2-oxohydro- pyrazinyl]butanamide 914-{5-(4-aminopiperidyl)-2-[6-(dimethyl- Aamino)(3-pyridyl)]-1-methyl-6-oxo(3-hydropyridyl)}-2-fluorobenzenecarbonitrile 924-{2-[6-(dimethylamino)(3-pyridyl)]-1- Amethyl-5-[4-(methylamino)piperidyl]-6-oxo-3-hydropyridyl}benzenecarbonitrile 934-[6-(4-aminopiperidin-1-yl)-4-methyl-3-(4- Cmethylphenyl)-5-oxopyrazin-2-yl]benzamide 945-(4-acetylphenyl)-3-(4-aminopiperidin-1-yl)-1- Cmethyl-6-(4-methylphenyl)pyrazin-2-one 953-(4-aminopiperidin-1-yl)-5-[4-(hydroxymeth- Cyl)phenyl]-1-methyl-6-(4-methylphenyl)pyrazin- 2-one 963-(4-aminopiperidin-1-yl)-1-methyl-6-(4-methyl- Bphenyl)-5-[4-(trifluoromethyl)phenyl]pyrazin- 2-one 973-(4-aminopiperidin-1-yl)-5-[4-(difluorometh- Byl)phenyl]-1-methyl-6-(4-methylphenyl)pyrazin- 2-one Note: Biochemicalassay IC₅₀ data are designated within the following ranges: A: ≦0.10 μMB: >0.10 μM to ≦1.0 μM C: >1.0 μM to ≦10 μM D: >10 μM

Example 2: In Vitro Enzyme Inhibition Assay—MAO Selectivity

Human recombinant monoamine oxidase proteins MAO-A and MAO-B wereobtained. MAOs catalyze the oxidative deamination of primary, secondaryand tertiary amines. In order to monitor MAO enzymatic activities and/ortheir inhibition rate by inhibitor(s) of interest, a fluorescent-based(inhibitor)-screening assay was performed.3-(2-Aminophenyl)-3-oxopropanamine (kynuramine dihydrobromide, SigmaAldrich), a non-fluorescent compound was chosen as a substrate.Kynuramine is a non-specific substrate for both MAOs activities. Whileundergoing oxidative deamination by MAO activities, kynuramine isconverted into 4-hydroxyquinoline (4-HQ), a resulting fluorescentproduct.

The monoamine oxidase activity was estimated by measuring the conversionof kynuramine into 4-hydroxyquinoline. Assays were conducted in 96-wellblack plates with clear bottom (Corning) in a final volume of 100 μl.The assay buffer was 100 mM HEPES, pH 7.5. Each experiment was performedin triplicate within the same experiment.

Briefly, a fixed amount of MAO (0.25 μg for MAO-A and 0.5 μg for AO-B)was incubated on ice for 15 minutes in the reaction buffer, in theabsence and/or in the presence of various concentrations of compounds asdisclosed herein (e.g., from 0 to 50 μM, depending on the inhibitorstrength). Tranylcypromine (Biomol International) was used as a controlfor inhibition.

After leaving the enzyme(s) interacting with the test compound, 60 to 90μM of kynuramine was added to each reaction for MAO-B and MAO-A assayrespectively, and the reaction was left for 1 hour at 37° C. in thedark. The oxidative deamination of the substrate was stopped by adding50 μl of 2N NaOH. The conversion of kynuramine to 4-hydroxyquinoline,was monitored by fluorescence (excitation at 320 nm, emission at 360 nm)using a microplate reader (Infinite 200, Tecan). Arbitrary units wereused to measure levels of fluorescence produced in the absence and/or inthe presence of test compound.

The maximum of oxidative deamination activity was obtained by measuringthe amount of 4-hydroxyquinoline formed from kynuramine deamination inthe absence of test compound and corrected for background fluorescence.The Ki (IC₅₀) of each inhibitor was determined at Vmax/2.

Example 3: LSD1 CD11b Cellular Assay

To analyze LSD1 inhibitor efficacy in cells, a CD11b flow cytometryassay was performed. LSD1 inhibition induces CD11b expression in THP-1(AML) cells which can be measured by flow cytometry. THP-1 cells wereseeded at 100,000 cells/well in 10% Fetal Bovine Serum containing RPMI1640 media in a 24 well plate with a final volume of 500 μL per well.LSD1 test compounds were serially diluted in DMSO. The dilutions wereadded to each well accordingly to a final concentration of 0.2% DMSO.The cells were incubated at 37 degrees Celsius in 5% CO₂ for 4 days. 250μL of each well was transferred to a well in a 96 well round bottomplate. The plate was centrifuged at 1200 rpm at 4 degrees Celsius in aBeckman Coulter Alegra 6KR centrifuge for 5 minutes. The media wasremoved leaving the cells at the bottom of the wells. The cells werewashed in 100 μL cold HBSS (Hank's Balanced Salt Solution) plus 2% BSA(Bovine Serum Albumin) solution and centrifuged at 1200 rpm at 4 degreesCelsius for 5 minutes. The wash was removed. The cells were resuspendedin 100 μL HBSS plus 2% BSA containing 1:15 dilution of APC conjugatedmouse anti-CD11b antibody (BD Pharmingen Cat#555751) and incubated onice for 25 minutes. The cells were centrifuged and washed two times in100 μl HBSS plus 2% BSA. After the final spin the cells were resuspendedin 100 μL HBSS plus 2% BSA containing 1 ug/mL DAPI(4′,6-diamidino-2-phenylindole). The cells were then analyzed by flowcytometry in a BD FACSAria machine. Cells were analyzed for CD11bexpression. The percent of CD11b expressing cells for each inhibitorconcentration was used to determine an IC₅₀ curve for each compoundanalyzed.

Table 4 provides the cellular IC₅₀ values of various substitutedheterocyclic compounds disclosed herein.

TABLE 4 Chemical THP-1 Synthesis IC₅₀ Example Name (μM) 14-[6-(4-aminopiperidin-1-yl)-4-benzyl-5-oxo-4,5- Bdihydropyrazin-2-yl]benzonitrile 2 4-[6-(4-aminopiperidin-1-yl)-4-[(4- Bmethylphenyl)methyl]-5-oxo-4,5-dihydropyrazin- 2-yl]benzonitrile 34-[6-(4-aminopiperidin-1-yl)-4-[(2- Bfluorophenyl)methyl]-5-oxo-4,5-dihydropyrazin- 2-yl]benzonitrile 44-[6-(4-aminopiperidin-1-yl)-4-(3-chloro-benzyl)- C5-oxo-4,5-dihydro-pyrazin-2-yl]-benzonitrile 54-[6-(4-amino-piperidin-1-yl)-4-methyl-5-oxo-4,5- Cdihydro-pyrazin-2-yl]-2-fluoro-benzonitrile 64-[6-(4-Amino-piperidin-1-yl)-4-ethyl-5-oxo-4,5- Cdihydro-pyrazin-2-yl]-2-fluoro-benzonitrile 74-[6-(4-amino-piperidin-1-yl)-4- Ccyclopropylmethyl-5-oxo-4,5-dihydro-pyrazin-2- yl]-2-fluoro-benzonitrile8 4-[6-(4-aminopiperidin-1-yl)-4-[(3- Bfluorophenyl)methyl]-5-oxo-4,5-dihydropyrazin- 2-yl]benzonitrile 94-[6-(4-aminopiperidin-1-yl)-4-[(4- Bfluorophenyl)methyl]-5-oxo-4,5-dihydropyrazin- 2-yl]benzonitrile 104-[6-(4-aminopiperidin-1-yl)-4-[(3- B methoxyphenyl)methyl]-5-oxo-4,5-dihydropyrazin-2-yl]benzonitrile 11 4-[6-(4-aminopiperidin-1-yl)-4-[(4-B methoxyphenyl)methyl]-5-oxo-4,5- dihydropyrazin-2-yl]benzonitrile 124-[6-(4-aminopiperidin-1-yl)-5-oxo-4-[(1R)-1- Cphenylethyl]-4,5-dihydropyrazin-2-yl]benzonitrile 134-[6-(4-aminopiperidin-1-yl)-5-oxo-4-[(1S)-1- Cphenylethyl]-4,5-dihydropyrazin-2-yl]benzonitrile 144-[6-(4-amino-piperidin-1-yl)-5-oxo-4- B(tetrahydro-furan-3-ylmethyl)-4,5-dihydro-pyrazin-2-yl]-2-fluoro-benzonitrile 154-[6-(4-amino-piperidin-1-yl)-5-oxo-4- B(tetrahydro-pyran-4-ylmethyl)-4,5-dihydro-pyrazin-2-yl]-2-fluoro-benzonitrile 164-[6-(4-aminopiperidin-1-yl)-4-methyl-3-(4- Amethylphenyl)-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 174-[4-methyl-3-(4-methylphenyl)-5-oxo-6-{[(3S)- Apyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin- 2-yl]benzonitrile 184-[4-methyl-3-(4-methylphenyl)-5-oxo-6-{[(3R)- Apyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin- 2-yl]benzonitrile 195-[6-(4-aminopiperidin-1-yl)-4-methyl-3-(4- Amethylphenyl)-5-oxo-4,5-dihydropyrazin-2- yl]pyridine-2-carbonitrile 204-{4-methyl-6-[4-(methylamino)piperidin-1-yl]-3- A(4-methylphenyl)-5-oxo-4,5-dihydropyrazin-2- yl}benzonitrile 214-[6-(4-amino-4-methylpiperidin-1-yl)-4-methyl- B3-(4-methylphenyl)-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 224-[6-(3-aminopiperidin-1-yl)-4-methyl-3-(4- Amethylphenyl)-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 234-[4-methyl-3-(4-methylphenyl)-6-{octahydro- A1H-pyrrolo[3,4-c]pyridin-5-yl}-5-oxo-4,5-dihydropyrazin-2-yl]benzonitrile 244-(6-{2,8-diazaspiro[4.5]decan-8-yl}-4-methyl-3- A(4-methylphenyl)-5-oxo-4,5-dihydropyrazin-2- yl)benzonitrile 254-(6-{decahydropyrrolo[3,4-d]azepin-6-yl}-4- Amethyl-3-(4-methylphenyl)-5-oxo-4,5- dihydropyrazin-2-yl)benzonitrile 264-[6-(4-aminopiperidin-1-yl)-4-ethyl-3-(4- Amethylphenyl)-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 274-[6-(4-aminopiperidin-1-yl)-3-(4-ethylphenyl)-4- Amethyl-5-oxo-4,5-dihydropyrazin-2- yl]benzonitrile 284-[3-(4-ethylphenyl)-4-methyl-5-oxo-6-{[(3S)- Bpyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin- 2-yl]benzonitrile 294-[3-(4-ethylphenyl)-4-methyl-5-oxo-6-{[(3R)- Bpyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin- 2-yl]benzonitrile 304-[6-(4-aminopiperidin-1-yl)-3-(4- A methoxyphenyl)-4-methyl-5-oxo-4,5-dihydropyrazin-2-yl]benzonitrile 314-[3-(4-methoxyphenyl)-4-methyl-5-oxo-6-{[(3S)- Apyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin- 2-yl]benzonitrile 324-[3-(4-methoxyphenyl)-4-methyl-5-oxo-6- B{[(3R)-pyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin-2-yl]benzonitrile 33 4-[6-(4-aminopiperidin-1-yl)-3-(4- Acyclopropylphenyl)-4-methyl-5-oxo-4,5- dihydropyrazin-2-yl]benzonitrile34 4-[3-(4-cyclopropylphenyl)-4-methyl-5-oxo-6- B{[(3S)-pyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin-2-yl]benzonitrile 354-[3-(4-cyclopropylphenyl)-4-methyl-5-oxo-6- B{[(3R)-pyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin-2-yl]benzonitrile 364-[6-(4-amino-piperidin-1-yl)-4-(2-methoxy- Aethyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]- benzonitrile 374-[6-(4-amino-piperidin-1-yl)-4-(2-hydroxy- Aethyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]- benzonitrile 384-[6-(4-amino-piperidin-1-yl)-3-(4-cyclopropyl- Aphenyl)-4-(2-hydroxy-ethyl)-5-oxo-4,5-dihydro-pyrazin-2-yl]-benzonitrile 394-[6-(4-amino-piperidin-1-yl)-4-(3-methoxy- Apropyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]- benzonitrile 404-[6-(4-amino-piperidin-1-yl)-4-(3-hydroxy- Apropyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]- benzonitrile 414-[6-(4-amino-piperidin-1-yl)-4-(2-methoxy- Aethyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]- 2-fluoro-benzonitrile42 4-[6-(4-amino-piperidin-1-yl)-4-(2-hydroxy- Aethyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]- 2-fluoro-benzonitrile43 4-(6-{[((3S)-pyrrolidin-3-yl)methyl]amino}-4-(2- Amethoxyethyl)-3-(4-methylphenyl)-5-oxo(4-hydropyrazin-2-yl))-2-fluorobenzenecarbonitrile 444-(6-{[((3S)-pyrrolidin-3-yl)methyl]amino}-4-(2- Ahydroxyethyl)-3-(4-methylphenyl)-5-oxo(4-hydropyrazin-2-yl))-2-fluorobenzenecarbonitrile 454-[6-(4-aminopiperidyl)-4-(3-methoxypropyl)-3- A(4-methylphenyl)-5-oxo(4-hydropyrazin-2-yl)]-2-fluorobenzenecarbonitrile 464-[6-(4-aminopiperidyl)-4-(3-hydroxypropyl)-3- A(4-methylphenyl)-5-oxo(4-hydropyrazin-2-yl)]-2-fluorobenzenecarbonitrile 474-[6-(4-amino-piperidin-1-yl)-4-methyl-5-oxo-3- Ap-tolyl-4,5-dihydro-pyrazin-2-yl]-2-fluoro- benzonitrile 484-[6-(4-amino-piperidin-1-yl)-4-methyl-5-oxo-3- Ap-tolyl-4,5-dihydro-pyrazin-2-yl]-3-fluoro- benzonitrile 494-[6-(4-amino-piperidin-1-yl)-4-methyl-3-(6- Amethyl-pyridin-3-yl)-5-oxo-4,5-dihydro-pyrazin- 2-yl]-benzonitrile 504-[6-(4-amino-piperidin-1-yl)-3-(4-cyclopropyl- Aphenyl)-4-methyl-5-oxo-4,5-dihydro-pyrazin-2- yl]-2-fluoro-benzonitrile51 5-[6-(4-amino-piperidin-1-yl)-3-(4-methoxy- Aphenyl)-4-methyl-5-oxo-4,5-dihydro-pyrazin-2-yl]-pyridine-2-carbonitrile 524-[6-(4-amino-piperidin-1-yl)-3-(4-methoxy- Aphenyl)-4-methyl-5-oxo-4,5-dihydro-pyrazin-2- yl]-2-fluoro-benzonitrile53 5-[6-(4-amino-piperidin-1-yl)-3-(4-cyclopropyl- Aphenyl)-4-methyl-5-oxo-4,5-dihydro-pyrazin-2-yl]-pyridine-2-carbonitrile 54 5-{3-(4-methoxy-phenyl)-4-methyl-5-oxo-6-A [((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro-pyrazin-2-yl}-pyridine-2-carbonitrile 552-fluoro-4-(3-(4-methoxy-phenyl)-4-methyl-5- Aoxo-6-[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro-pyrazin-2-yl}-benzonitrile 565-{3-(4-ethyl-phenyl)-4-methyl-5-oxo-6-[((3S)- Apyrrolidin-3-ylmethyl)-amino]-4,5-dihydro-pyrazin-2-yl}-pyridine-2-carbonitrile 574-{3-(4-ethyl-phenyl)-4-methyl-5-oxo-6-[((3S)- Apyrrolidin-3-ylmethyl)-amino]-4,5-dihydro-pyrazin-2-yl}-2-fluoro-benzonitrile 585-{3-(4-cyclopropyl-phenyl)-4-methyl-5-oxo-6- B[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro-pyrazin-2-yl}-pyridine-2-carbonitrile 594-{3-(4-cyclopropyl-phenyl)-4-methyl-5-oxo-6- A[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro-pyrazin-2-yl}-2-fluoro-benzonitrile 604-[6-(4-amino-piperidin-1-yl)-4-methyl-3-(1- Amethyl-1H-benzoimidazol-5-yl)-5-oxo-4,5-dihydro-pyrazin-2-yl]-2-fluoro-benzonitrile 614-{3-(4-cyclopropylmethoxy-phenyl)-4-methyl-5- Aoxo-6-[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro-pyrazin-2-yl}-2-fluoro-benzonitrile 624-{3-(4-ethoxy-phenyl)-4-methyl-5-oxo-6-[((3S)- Apyrrolidin-3-ylmethyl)-amino]-4,5-dihydro-pyrazin-2-yl}-2-fluoro-benzonitrile 632-fluoro-4-[4-methyl-5-oxo-6-[((3S)-pyrrolidin-3- Aylmethyl)-amino]-3-(4-trifluoromethoxy-phenyl)-4,5-dihydro-pyrazin-2-yl]-benzonitrile 642-Fluoro-4-[4-(3-hydroxy-propyl)-6-(4- Amethylamino-piperidin-1-yl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]-benzonitrile 652-Fluoro-4-[4-(2-hydroxy-ethyl)-6-(4- Amethylamino-piperidin-1-yl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2-yl]-benzonitrile 664-[6-(4-Amino-piperidin-1-yl)-4-methyl-3-(6- Amethyl-pyridin-3-yl)-5-oxo-4,5-dihydro-pyrazin-2-yl]-2-fluoro-benzonitrile 67 4-(5-(4-aminopiperidin-1-yl)-2-(4- Amethoxyphenyl)-1-methyl-6-oxo-1,6-dihydropyridin-3-yl)-2-fluorobenzonitrile 685-(5-(4-aminopiperidin-1-yl)-2-(4- B methoxyphenyl)-1-methyl-6-oxo-1,6-dihydropyridin-3-yl)picolinonitrile 69(S)-5-(2-(4-methoxyphenyl)-1-methyl-6-oxo-5- B(pyrrolidin-3-ylmethylamino)-1,6-dihydropyridin- 3-yl)picolinonitrile 70(S)-2-fluoro-4-(2-(4-methoxyphenyl)-1-methyl-6- Aoxo-5-(pyrrolidin-3-ylmethylamino)-1,6- dihydropyridin-3-yl)benzonitrile71 (S)-5-(2-(4-ethylphenyl)-1-methyl-6-oxo-5- A(pyrrolidin-3-ylmethylamino)-1,6-dihydropyridin- 3-yl)picolinonitrile 72(S)-4-(2-(4-ethylphenyl)-1-methyl-6-oxo-5- A(pyrrolidin-3-ylmethylamino)-1,6-dihydropyridin-3-yl)-2-fluorobenzonitrile 73(S)-5-(2-(4-cyclopropylphenyl)-1-methyl-6-oxo-5- A(pyrrolidin-3-ylmethylamino)-1,6-dihydropyridin- 3-yl)picolinonitrile 74(S)-4-(2-(4-cyclopropylphenyl)-1-methyl-6-oxo-5- A(pyrrolidin-3-ylmethylamino)-1,6-dihydropyridin-3-yl)-2-fluorobenzonitrile 754-[5-(4-aminopiperidin-1-yl)-1-methyl-2-(4- Amethylphenyl)-6-oxopyridin-3-yl]benzonitrile 764-[5-(4-aminopiperidin-1-yl)-1-methyl-2-(4- Amethylphenyl)-6-oxopyridin-3-yl]-2- fluorobenzonitrile 774-[5-(4-aminopiperidin-1-yl)-2-(4- Amethoxyphenyl)-1-methyl-6-oxopyridin-3- yl]benzonitrile 784-{2-(4-methoxyphenyl)-1-methyl-5-[4- A (methylamino)piperidyl]-6-oxo-3-hydropyridyl}benzenecarbonitrile 792-fluoro-4-{2-(4-methoxyphenyl)-1-methyl-5-[4- A(methylamino)piperidyl]-6-oxo(3- hydropyridyl)}benzenecarbonitrile 804-{1-methyl-5-[4-(methylamino)piperidyl]-2-(4- A methylphenyl)-6-oxo-3-hydropyridyl}benzenecarbonitrile 81 4-[5-(4-aminopiperidin-1-yl)-1- A(cyclopropylmethyl)-2-(4-methoxyphenyl)-6- oxopyridin-3-yl]benzonitrile82 4-[5-(4-aminopiperidin-1-yl)-1-ethyl-2-(4- Amethoxyphenyl)-6-oxopyridin-3-yl]benzonitrile 834-[5-(4-aminopiperidin-1-yl)-2-(3-fluoro-4- Amethoxyphenyl)-1-methyl-6-oxopyridin-3- yl]benzonitrile 844-[5-(4-aminopiperidin-1-yl)-2-(3-fluoro-4- Amethoxyphenyl)-1-methyl-6-oxopyridin-3-yl]-2- fluorobenzonitrile 854-[5-(4-aminopiperidin-1-yl)-1- A(cyclopropylmethyl)-2-(4-methoxyphenyl)-6-oxopyridin-3-yl]-2-fluorobenzonitrile 862-fluoro-4-[2-(3-fluoro-4-methoxyphenyl)-1- Amethyl-5-[4-(methylamino)piperidin-1-yl]-6- oxopyridin-3-yl]benzonitrile87 4-[5-(4-aminopiperidin-1-yl)-1-ethyl-2-(4- Amethoxyphenyl)-6-oxopyridin-3-yl]-2- fluorobenzonitrile 882-[3-(4-aminopiperidyl)-5-(4-cyano-3- Afluorophenyl)-6-(4-methoxyphenyl)-2- oxohydropyrazinyl]acetamide 893-[3-(4-aminopiperidyl)-5-(4-cyano-3- Afluorophenyl)-6-(4-methoxyphenyl)-2- oxohydropyrazinyl]propanamide 904-[3-(4-aminopiperidyl)-5-(4-cyano-3- Afluorophenyl)-6-(4-methoxyphenyl)-2- oxohydropyrazinyl]butanamide 914-{5-(4-aminopiperidyl)-2-[6-(dimethylamino)(3- Apyridyl)]-1-methyl-6-oxo(3-hydropyridyl)}-2- fluorobenzenecarbonitrile92 4-{2-[6-(dimethylamino)(3-pyridyl)]-1-methyl-5- A[4-(methylamino)piperidyl]-6-oxo-3- hydropyridyl}benzenecarbonitrileNote: Cellular assay IC₅₀ data are designated within the followingranges: A: ≦0.10 μM B: >0.10 μM to ≦1.0 μM C: >1.0 μM to ≦10 μM D: >10μM

Example 4: Kasumi-1 AML Cell Line Proliferation Assay (Cell-MTS Assay)

Colorimetric cellular assay to assess the ability of LSD-1 smallmolecule inhibitors to effect the proliferation of the established AMLcancer cell line Kasumi-1.

Assay Background

The LSD-1 protein has been shown to play a key role in the biology of avariety of cancer types including SCLC and AML. To demonstrate smallmolecule inhibition of LSD-1 as a potential anti-cancer therapy, anassay to measure the degree of proliferative inhibition in anestablished cancer cell line of AML was implemented.

Assay Principle

This Cell-MTS assay is a 7-day plate based colorimetric assay whichquantifies the amount of newly generated NADH in the presence andabsence of test compound. These NADH levels are used as a proxy for thequantification of cancer cell proliferation.

Assay Method in Brief

The established cancer cell line Kasumi-1 with a verified p53 mutationwere purchased from American Type Culture Collection (ATCC) androutinely passaged according to ATCC published protocols. For routineassay these cells were seeded at a density of 20,000 cells per 96-well.24 hours after plating, cells received an 11 point dilution of testcompound with final concentration ranges from 100 μM to 2.0 nM. Cellsare incubated in the presence of compound for 168 hours at 37° C., 5%CO₂. At the end of this compound incubation period, 80 μl of media isremoved and 20 μL of CellTiter 96® AQueous Non-Radioactive CellProliferation Assay solution (Promega) is added. The cells are incubateduntil the OD490 is >0.6. IC₅₀ values are calculated using the IDBS XLfitsoftware package and include background subtracted OD490 values andnormalization to DMSO controls.

Table 5 provides the Kasumi-1 cellular IC₅₀ values of varioussubstituted heterocyclic compounds disclosed herein.

TABLE 5 Chemical Kasumi-1 Synthesis IC₅₀ Example Name (μM) 284-[3-(4-ethylphenyl)-4-methyl-5-oxo-6- A{[(3S)-pyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin-2-yl]benzonitrile 314-[3-(4-methoxyphenyl)-4-methyl-5-oxo-6- A{[(3S)-pyrrolidin-3-ylmethyl]amino}-4,5-dihydropyrazin-2-yl]benzonitrile 334-[6-(4-aminopiperidin-1-yl)-3-(4- Acyclopropylphenyl)-4-methyl-5-oxo-4,5- dihydropyrazin-2-yl]benzonitrile36 4-[6-(4-amino-piperidin-1-yl)-4-(2-methoxy- Aethyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2- yl]-benzonitrile 404-[6-(4-amino-piperidin-1-yl)-4-(3-hydroxy- Apropyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2- yl]-benzonitrile 424-[6-(4-amino-piperidin-1-yl)-4-(2-hydroxy- Aethyl)-5-oxo-3-p-tolyl-4,5-dihydro-pyrazin-2- yl]-2-fluoro-benzonitrile44 4-(6-{[((3S)-pyrrolidin-3-yl)methyl]amino}- A4-(2-hydroxyethyl)-3-(4-methylphenyl)-5-oxo(4-hydropyrazin-2-yl))-2-fluorobenzenecarbonitrile 464-[6-(4-aminopiperidyl)-4-(3-hydroxypropyl)-3- A(4-methylphenyl)-5-oxo(4-hydropyrazin-2-yl)]-2-fluorobenzenecarbonitrile 552-fluoro-4-{3-(4-methoxy-phenyl)-4-methyl-5- Aoxo-6-[((3S)-pyrrolidin-3-ylmethyl)-amino]-4,5-dihydro-pyrazin-2-yl}-benzonitrile 604-[6-(4-amino-piperidin-1-yl)-4-methyl-3-(1- Amethyl-1H-benzoimidazol-5-yl)-5-oxo-4,5-dihydro-pyrazin-2-yl]-2-fluoro-benzonitrile 664-[6-(4-Amino-piperidin-1-yl)-4-methyl-3-(6- Amethyl-pyridin-3-yl)-5-oxo-4,5-dihydro-pyrazin-2-yl]-2-fluoro-benzonitrile 70(S)-2-fluoro-4-(2-(4-methoxyphenyl)-1-methyl-6- Aoxo-5-(pyrrolidin-3-ylmethylamino)-1,6- dihydropyridin-3-yl)benzonitrile74 (S)-4-(2-(4-cyclopropylphenyl)-1-methyl-6-oxo-5- A(pyrrolidin-3-ylmethylamino)-1,6-dihydropyridin-3-yl)-2-fluorobenzonitrile 754-[5-(4-aminopiperidin-1-yl)-1-methyl-2-(4- Amethylphenyl)-6-oxopyridin-3-yl]benzonitrile 764-[5-(4-aminopiperidin-1-yl)-1-methyl-2-(4- Amethylphenyl)-6-oxopyridin-3-yl]-2- fluorobenzonitrile 774-[5-(4-aminopiperidin-1-yl)-2-(4- Amethoxyphenyl)-1-methyl-6-oxopyridin-3- yl]benzonitrile 834-[5-(4-aminopiperidin-1-yl)-2-(3-fluoro-4- Amethoxyphenyl)-1-methyl-6-oxopyridin-3- yl]benzonitrile 844-[5-(4-aminopiperidin-1-yl)-2-(3-fluoro-4- Amethoxyphenyl)-1-methyl-6-oxopyridin-3-yl]- 2-fluorobenzonitrile 914-{5-(4-aminopiperidyl)-2-[6-(dimethyl- Aamino)(3-pyridyl)]-1-methyl-6-oxo(3-hydro-pyridyl)}-2-fluorobenzenecarbonitrile Note: Cellular assay IC₅₀ data aredesignated within the following ranges: A: ≦0.10 μM B: >0.10 μM to ≦1.0μM C: >1.0 μM to ≦10 μM D: >10 μM

Example 5a: In Vivo Xenograph Study—MCF-7 Xenograph

Time release pellets containing 0.72 mg 17-β Estradiol aresubcutaneously implanted into nu/nu mice. MCF-7 cells are grown in RPMIcontaining 10% FBS at 5% CO₂, 37° C. Cells are spun down andre-suspended in 50% RPMI (serum free) and 50% Matrigel at 1×10⁷cells/mL. MCF-7 cells are subcutaneously injected (100 μL/animal) on theright flank 2-3 days post pellet implantation and tumor volume(length×width²/2) is monitored bi-weekly. When tumors reach an averagevolume of ˜200 mm³ animals are randomized and treatment is started.Animals are treated with vehicle or compound daily for 4 weeks. Tumorvolume and body weight are monitored bi-weekly throughout the study. Atthe conclusion of the treatment period, plasma and tumor samples aretaken for pharmacokinetic and pharmacodynamic analyses, respectively.

Example 5b: In Vivo Xenograph Study—LNCaP Xenograph

LNCaP cells with a stable knockdown of LSD1 (shLSD1 cells) or controlcells (such as shNTC cells) are inoculated in the dorsal flank of nudemice by subcutaneous injection (such as 3×10⁶ cells in 100 μl of 50%RPMI 1640/BD Matrigel). Mouse weight and tumor size are measured onceper week and tumor volume is estimated using the formula (7i/6)(L×W),where L=length of tumor and W=width of tumor. A two sample t-test isperformed to determine statistical differences in mean tumor volumebetween the two groups.

Unmodified LNCaP cells are inoculated by subcutaneous injection into thedorsal flank of nude mice (such as 3×10⁶ cells in 100 μl of 50% RPMI1640/BD Matrigel). After three weeks, mice are injectedintraperitoneally once per day with water (control), pargyline (0.53 mgor 1.59 mg; 1 or 3 mM final concentration, assuming 70%bioavailability), or XB154 (4 or 20 μg; 1 or 5 μM final concentration,assuming 70% bioavailability) or treated with a test compound (5 mg/kgeach week or 10 mg/kg each week). Treatment continues for three weeks,during which time mouse weight and tumor volume are measured as above.

shLSD1 LNCaP cells or control cells are injected in nude mice as above.After three weeks, mice are treated with 2.6 μg mitomycin C (predictedfinal concentration of 1 μM assuming 40% bioavailability), olaparib (forexample, about 0.5 mg/kg to 25 mg/kg), or vehicle intraperitoneally onceper day for three weeks. In other examples, unmodified LNCaP cells areinjected in nude mice as above.

After three weeks, mice are treated with test compounds, or vehicle asabove, plus MMC or olaparib. Treatment continues for three weeks, duringwhich time mouse weight and tumor volume are measured as above.

A decrease in tumor volume compared to control in mice injected withshLSD1 cells indicates that LSD1 inhibition decreases tumor growth invivo.

Similarly, a decrease in tumor volume compared to control in miceinjected with LNCaP cells and treated with a compound disclosed hereinindicates that LSD1 inhibition decreases tumor growth in vivo. Finally,a decrease in tumor volume in mice injected with LNCaP cells and treatedwith a compound disclosed herein plus olaparib as compared to micetreated with a compound disclosed herein alone indicates that inhibitionof LSD1 plus inhibition of PARP decreases tumor growth in vivo.

The harvested xenograft tissue is examined for evidence of LSD1inhibition. This is assessed with Western blots to examine global levelsof the 2MK4 and 2MK9 histone marks, expression of FA/BRCA genes, FANCD2ubiquitination, and LSD1 protein levels in the cases of the shRNA cells.A decrease in one or more of these parameters indicates the effectiveinhibition of LSD 1. Additionally, effects on DNA damage repair areassessed with staining for H2AX foci.

III. Preparation of Pharmaceutical Dosage Forms Example 1: Oral Tablet

A tablet is prepared by mixing 48% by weight of a compound of Formula(I) or a pharmaceutically acceptable salt thereof, 45% by weight ofmicrocrystalline cellulose, 5% by weight of low-substitutedhydroxypropyl cellulose, and 2% by weight of magnesium stearate. Tabletsare prepared by direct compression. The total weight of the compressedtablets is maintained at 250-500 mg.

We claim:
 1. A compound having the structure of Formula (I), or apharmaceutically acceptable salt thereof,

wherein, A is N or C—R, wherein R is hydrogen or optionally substitutedalkyl; W¹ and W² are independently chosen from N, C—H, or C—F; X ishydrogen, halogen, optionally substituted alkyl, optionally substitutedcycloalkylalkyl, optionally substituted heterocyclylalkyl, optionallysubstituted aralkyl, optionally substituted heteroarylalkyl, optionallysubstituted alkyne, optionally substituted carbocyclylalkynyl,optionally substituted aryl, or optionally substituted heteroaryl; Y isoptionally substituted alkyl, optionally substituted cycloalkylalkyl,heterocyclylalkyl or aralkyl; and Z is an optionally substituted groupchosen from N-heterocyclyl, —O-heterocyclylalkyl,—N(H)-heterocyclylalkyl, or —N(Me)-heterocyclylalkyl.
 2. The compound orpharmaceutically acceptable salt of claim 1, wherein W² is C—H.
 3. Thecompound or pharmaceutically acceptable salt of claim 1 or 2, wherein W¹is C—F.
 4. The compound or pharmaceutically acceptable salt of claim 1or 2, wherein W¹ is C—H.
 5. The compound or pharmaceutically acceptablesalt of claim 1 or 2, wherein W¹ is N.
 6. The compound orpharmaceutically acceptable salt of claims 1-5, wherein X is hydrogen.7. The compound or pharmaceutically acceptable salt of claims 1-5,wherein X is optionally substituted alkyl.
 8. The compound orpharmaceutically acceptable salt of claims 1-5, wherein X is optionallysubstituted alkyne.
 9. The compound or pharmaceutically acceptable saltof claim 1-5, wherein X is optionally substituted carbocyclylalkynyl.10. The compound or pharmaceutically acceptable salt of claims 1-5,wherein X is optionally substituted aryl, or optionally substitutedheteroaryl.
 11. The compound or pharmaceutically acceptable salt ofclaims 1-5, wherein X is optionally substituted aryl.
 12. The compoundor pharmaceutically acceptable salt of claim 11, wherein the optionallysubstituted aryl is an optionally substituted phenyl.
 13. The compoundor pharmaceutically acceptable salt of claims 1-5, wherein X isoptionally substituted heteroaryl.
 14. The compound or pharmaceuticallyacceptable salt of claim 13, wherein the optionally substitutedheteroaryl is chosen from an optionally substituted pyridinyl,optionally substituted pyrazolyl, or optionally substituted indazolyl.15. The compound or pharmaceutically acceptable salt of claims 1-14,wherein Z is an optionally substituted —O-heterocyclylalkyl.
 16. Thecompound or pharmaceutically acceptable salt of claims 1-14, wherein Zis an optionally substituted —N(H)-heterocyclylalkyl.
 17. The compoundor pharmaceutically acceptable salt of claims 1-14, wherein Z is anoptionally substituted —N(Me)-heterocyclylalkyl.
 18. The compound orpharmaceutically acceptable salt of claims 15-19, wherein theheterocyclylalkyl group has the formula —R^(c)-heterocyclyl and theR^(c) is an optionally substituted C₁-C₃ alkylene chain.
 19. Thecompound or pharmaceutically acceptable salt of claims 15-17, whereinthe heterocyclylalkyl group has the formula —R^(c)-heterocyclyl and theR^(c) is an optionally substituted C₁ alkylene chain.
 20. The compoundor pharmaceutically acceptable salt of claims 15-17, wherein theheterocyclylalkyl group has the formula —R^(c)-heterocyclyl and theheterocyclyl is an optionally substituted nitrogen-containing 4-, 5-,6-, or 7-membered heterocyclyl.
 21. The compound or pharmaceuticallyacceptable salt of claims 1-14, wherein Z is an optionally substitutedN-heterocyclyl.
 22. The compound or pharmaceutically acceptable salt ofclaim 21, wherein the optionally substituted N-heterocyclyl is a 4-, 5-,6-, or 7-membered N-heterocyclyl.
 23. The compound or pharmaceuticallyacceptable salt of claim 21, wherein the optionally substitutedN-heterocyclyl is a 6-membered N-heterocyclyl.
 24. The compound orpharmaceutically acceptable salt of claim 21, wherein the optionallysubstituted N-heterocyclyl is an optionally substituted piperidine. 25.The compound or pharmaceutically acceptable salt of claim 24, whereinthe optionally substituted piperidine is an optionally substituted4-aminopiperidine.
 26. The compound or pharmaceutically acceptable saltof claims 1-25, wherein Y is optionally substituted cycloalkylalkyl. 27.The compound or pharmaceutically acceptable salt of claims 1-25, whereinY is optionally substituted alkyl.
 28. The compound or pharmaceuticallyacceptable salt of claims 1-25, wherein Y is optionally substitutedheterocyclylalkyl.
 29. The compound or pharmaceutically acceptable saltof claims 1-25, wherein Y is optionally substituted aralkyl.
 30. Thecompound or pharmaceutically acceptable salt of claim 27, wherein theoptionally substituted alkyl is an optionally substituted C₁-C₃ alkyl.31. The compound or pharmaceutically acceptable salt of claim 27,wherein the optionally substituted alkyl is an optionally substituted C₁alkyl.
 32. The compound or pharmaceutically acceptable salt of claim 27,wherein the optionally substituted alkyl is a methyl group.
 33. Thecompound or pharmaceutically acceptable salt of any of the proceedingclaims, wherein A is N.
 34. The compound or pharmaceutically acceptablesalt of any one of claims 1-32, wherein A is C—R.
 35. The compound orpharmaceutically acceptable salt of any one of claims 1-32, wherein A isC—R, and R is hydrogen
 36. The compound or pharmaceutically acceptablesalt of any one of claims 1-32, wherein A is C—R and R is optionallysubstituted alkyl.
 37. A pharmaceutical composition comprising acompound of Formula (I), or a pharmaceutically acceptable salt thereof,and a pharmaceutically acceptable excipient.
 38. A method of regulatinggene transcription in a cell comprising inhibiting lysine-specificdemethylase 1 activity by exposing the lysine-specific demethylase 1enzyme to a compound of Formula (I).
 39. A method of treating cancer ina patient in need thereof, comprising administering to the patient acompound of Formula (I), or a pharmaceutically acceptable salt thereof.